The cytoplasmic and nuclear proteasome complexes function to degrade ubiquitin-labeled proteins that are targeted for down regulation or disposal due to misfolding. Volume 4, Number 2 BIOFILES Life Science Protease Inhibition and Detection Broad Spectrum Protease Inhibitors Inhibitor Cocktails and Tablets Inhibitor Specificity Index for: Proteasome and Lysozomal Proteases Complement and Coagulation Factors Renin-Angiotensin System Enzymes Extracellular Matrix/MMPs Other Proteases Protease Detection: Kits and Substrates
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The cytoplasmic and nuclear proteasome complexes function to degrade ubiquitin-labeled proteins that are targeted for down regulation or disposal due to misfolding
Volume 4 Number 2
BioFilesLife Science
Protease Inhibition and Detection
Broad Spectrum Protease Inhibitors
Inhibitor Cocktails and Tablets
Inhibitor Specificity Index forProteasome and Lysozomal Proteases Complement and Coagulation FactorsRenin-Angiotensin System EnzymesExtracellular MatrixMMPsOther Proteases
Protease Detection Kits and Substrates
2
Life Science
BioFilesVolume 4 Number 2
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Life Science TVFeaturing the Non-Specific Protease Activity Assay
The Sigmareg Life Science TV Web site is home to instructional informational and protocol based videos With regular updates to the library you will find videos covering many topics across the life science disciplines Currently available videos include
bull ATP Synthase Animation
bull GenomePlexreg Tutorial
bull TargeTronreg Technical Animation
The latest video addition to Life Science TV is titledNon-Specific Protease Activity Assay
This video displays a detailed protocol for determining the activity of proteases The universal assay procedure is useful for experiments ranging from sample preparation to QC analysis
View all Sigma Life Science Videos at sigma-aldrichcomlsvideos
Table of Contents
Introduction 3
Broad Spectrum Inhibitors of Proteolytic Enzyme Classes 4
Proteolytic hydrolysis of peptide bonds was first studied as a function of digestion in higher mammals It is now recognized as an essential and ubiquitous mechanism for the regulation of a myriad of physiological processes
Inhibition of proteolytic activity is usually employed for two purposes
For the prevention of unwanted degradation of proteins during their isolation and characterization
To study the regulatory aspects of specific proteolytic events as they relate to cellular processes Some of the most studied proteolytic processes include blood coagulation and complement cascades hormonal regulation apoptosis extracellular matrix degradation proteasome and lysozomal regulation and disease states such as Alzheimerrsquos and viral replication
Four main classes of proteolyic enzymes have been routinely utilized to describe proteases The serine proteases are probably the best characterized This class of proteases includes trypsin chymotrypsin and elastase The cysteine protease class includes papain calpain and lysozomal cathepsins Aspartic proteases include pepsin and rennin Metalloproteinases include thermolysin and carboxypeptidase A
During isolation and characterization one or all four classes of proteases may pose a threat to the fate of a protein Broad-spectrum protease inhibitors and mixtures (or cocktails) have been developed to protect the integrity of isolated proteins Sigmareg offers and manufactures the broadest range of protease inhibitors and inhibitor cocktails of any supplier Sigma inhibitor cocktails have been specifically formulated for particular applications as they relate to the biological source or method of expression
Protease inhibitors can be added during cell growth and protein expression or can be added at the time of extraction
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Bro
ad S
pec
tru
m In
hib
ito
rs o
f Pr
ote
oly
tic
Enzy
me
Cla
sses
Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics
Typical Working Concentrations
Preparation of Stock Solutions
Serine Protease Inhibitors
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM 100 mM in water stable 1 month at -20 degC
A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC
D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC
T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl
T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Cysteine Protease Inhibitors
E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
Aspartic Protease Inhibitors
P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Metalloproteinase Inhibitors
ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water
P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC
R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase
1-10 mM 1 mM in water stable 1 month at -20 degC
B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data
Our Innovation Your Research mdash Shaping the Future of Life Science 5
Protease In
hib
itor Pan
el
Protease Inhibitor PanelProtease Inhibitor Panel
Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases
Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility
As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture
Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor
INHIB1-1KT 1 kit
INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility
Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location
Simple to use
Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)
capable of the cleavage
sigma-aldrichcom
Protease Finder
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Pro
teas
e In
hib
ito
r C
ock
tails
an
d T
able
ts
Protease Inhibitor Cocktails and TabletsYou need a cocktail
End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg
For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)
One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract
S8820-20TAB 20 tablets
Protease Inhibitor Cocktail
for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA
One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin
P2714-1BTL 1 bottle
for use in purification of Histidine-tagged proteins DMSO solution
Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon
One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml
P8849-1ML 1 mL
P8849-5ML 5 mL
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Our Innovation Your Research mdash Shaping the Future of Life Science 7
Protease In
hib
itor C
ocktails an
d Tab
lets
for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators
One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver
P8340-1ML 1 mL
P8340-5ML 5 mL
for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators
Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins
Solution in DMSO (D 2650 Hybri-Max)
P1860-1ML 1 mL
for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA
Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli
Supplied with a vial of DMSO
P8465-5ML 5 mL
P8465-25ML 25 mL
for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 20 g of yeast
P8215-1ML 1 mL
P8215-5ML 5 mL
for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml
Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
P9599-1ML 1 mL
P9599-5ML 5 mL
While Sigma scientists have invested
considerable resources to formulate
the most effective cocktails suitable
for as many applications as possible
each cell line extractionpurification
procedure and expression system poses a different
set of proteolytic challenges We would like to solicit
feedback from researchers who have encountered
problems with standard cocktail formulations as
well as those who have developed novel alternative
strategies for protease inhibition Visit our BioBlog at
sigmabioblogscom to learn more
8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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ng
s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
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Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
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Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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e D
etec
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
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to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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Life Science TVFeaturing the Non-Specific Protease Activity Assay
The Sigmareg Life Science TV Web site is home to instructional informational and protocol based videos With regular updates to the library you will find videos covering many topics across the life science disciplines Currently available videos include
bull ATP Synthase Animation
bull GenomePlexreg Tutorial
bull TargeTronreg Technical Animation
The latest video addition to Life Science TV is titledNon-Specific Protease Activity Assay
This video displays a detailed protocol for determining the activity of proteases The universal assay procedure is useful for experiments ranging from sample preparation to QC analysis
View all Sigma Life Science Videos at sigma-aldrichcomlsvideos
Table of Contents
Introduction 3
Broad Spectrum Inhibitors of Proteolytic Enzyme Classes 4
Proteolytic hydrolysis of peptide bonds was first studied as a function of digestion in higher mammals It is now recognized as an essential and ubiquitous mechanism for the regulation of a myriad of physiological processes
Inhibition of proteolytic activity is usually employed for two purposes
For the prevention of unwanted degradation of proteins during their isolation and characterization
To study the regulatory aspects of specific proteolytic events as they relate to cellular processes Some of the most studied proteolytic processes include blood coagulation and complement cascades hormonal regulation apoptosis extracellular matrix degradation proteasome and lysozomal regulation and disease states such as Alzheimerrsquos and viral replication
Four main classes of proteolyic enzymes have been routinely utilized to describe proteases The serine proteases are probably the best characterized This class of proteases includes trypsin chymotrypsin and elastase The cysteine protease class includes papain calpain and lysozomal cathepsins Aspartic proteases include pepsin and rennin Metalloproteinases include thermolysin and carboxypeptidase A
During isolation and characterization one or all four classes of proteases may pose a threat to the fate of a protein Broad-spectrum protease inhibitors and mixtures (or cocktails) have been developed to protect the integrity of isolated proteins Sigmareg offers and manufactures the broadest range of protease inhibitors and inhibitor cocktails of any supplier Sigma inhibitor cocktails have been specifically formulated for particular applications as they relate to the biological source or method of expression
Protease inhibitors can be added during cell growth and protein expression or can be added at the time of extraction
4 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Bro
ad S
pec
tru
m In
hib
ito
rs o
f Pr
ote
oly
tic
Enzy
me
Cla
sses
Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics
Typical Working Concentrations
Preparation of Stock Solutions
Serine Protease Inhibitors
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM 100 mM in water stable 1 month at -20 degC
A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC
D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC
T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl
T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Cysteine Protease Inhibitors
E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
Aspartic Protease Inhibitors
P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Metalloproteinase Inhibitors
ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water
P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC
R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase
1-10 mM 1 mM in water stable 1 month at -20 degC
B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data
Our Innovation Your Research mdash Shaping the Future of Life Science 5
Protease In
hib
itor Pan
el
Protease Inhibitor PanelProtease Inhibitor Panel
Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases
Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility
As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture
Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor
INHIB1-1KT 1 kit
INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility
Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location
Simple to use
Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)
capable of the cleavage
sigma-aldrichcom
Protease Finder
6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e In
hib
ito
r C
ock
tails
an
d T
able
ts
Protease Inhibitor Cocktails and TabletsYou need a cocktail
End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg
For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)
One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract
S8820-20TAB 20 tablets
Protease Inhibitor Cocktail
for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA
One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin
P2714-1BTL 1 bottle
for use in purification of Histidine-tagged proteins DMSO solution
Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon
One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml
P8849-1ML 1 mL
P8849-5ML 5 mL
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BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic
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Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you
Our Innovation Your Research mdash Shaping the Future of Life Science 7
Protease In
hib
itor C
ocktails an
d Tab
lets
for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators
One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver
P8340-1ML 1 mL
P8340-5ML 5 mL
for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators
Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins
Solution in DMSO (D 2650 Hybri-Max)
P1860-1ML 1 mL
for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA
Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli
Supplied with a vial of DMSO
P8465-5ML 5 mL
P8465-25ML 25 mL
for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 20 g of yeast
P8215-1ML 1 mL
P8215-5ML 5 mL
for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml
Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
P9599-1ML 1 mL
P9599-5ML 5 mL
While Sigma scientists have invested
considerable resources to formulate
the most effective cocktails suitable
for as many applications as possible
each cell line extractionpurification
procedure and expression system poses a different
set of proteolytic challenges We would like to solicit
feedback from researchers who have encountered
problems with standard cocktail formulations as
well as those who have developed novel alternative
strategies for protease inhibition Visit our BioBlog at
sigmabioblogscom to learn more
8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
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Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
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Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
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of continually curated biological data
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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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Proteolytic hydrolysis of peptide bonds was first studied as a function of digestion in higher mammals It is now recognized as an essential and ubiquitous mechanism for the regulation of a myriad of physiological processes
Inhibition of proteolytic activity is usually employed for two purposes
For the prevention of unwanted degradation of proteins during their isolation and characterization
To study the regulatory aspects of specific proteolytic events as they relate to cellular processes Some of the most studied proteolytic processes include blood coagulation and complement cascades hormonal regulation apoptosis extracellular matrix degradation proteasome and lysozomal regulation and disease states such as Alzheimerrsquos and viral replication
Four main classes of proteolyic enzymes have been routinely utilized to describe proteases The serine proteases are probably the best characterized This class of proteases includes trypsin chymotrypsin and elastase The cysteine protease class includes papain calpain and lysozomal cathepsins Aspartic proteases include pepsin and rennin Metalloproteinases include thermolysin and carboxypeptidase A
During isolation and characterization one or all four classes of proteases may pose a threat to the fate of a protein Broad-spectrum protease inhibitors and mixtures (or cocktails) have been developed to protect the integrity of isolated proteins Sigmareg offers and manufactures the broadest range of protease inhibitors and inhibitor cocktails of any supplier Sigma inhibitor cocktails have been specifically formulated for particular applications as they relate to the biological source or method of expression
Protease inhibitors can be added during cell growth and protein expression or can be added at the time of extraction
4 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Bro
ad S
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ito
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f Pr
ote
oly
tic
Enzy
me
Cla
sses
Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics
Typical Working Concentrations
Preparation of Stock Solutions
Serine Protease Inhibitors
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM 100 mM in water stable 1 month at -20 degC
A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC
D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC
T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl
T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Cysteine Protease Inhibitors
E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
Aspartic Protease Inhibitors
P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Metalloproteinase Inhibitors
ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water
P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC
R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase
1-10 mM 1 mM in water stable 1 month at -20 degC
B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data
Our Innovation Your Research mdash Shaping the Future of Life Science 5
Protease In
hib
itor Pan
el
Protease Inhibitor PanelProtease Inhibitor Panel
Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases
Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility
As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture
Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor
INHIB1-1KT 1 kit
INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility
Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location
Simple to use
Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)
capable of the cleavage
sigma-aldrichcom
Protease Finder
6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e In
hib
ito
r C
ock
tails
an
d T
able
ts
Protease Inhibitor Cocktails and TabletsYou need a cocktail
End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg
For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)
One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract
S8820-20TAB 20 tablets
Protease Inhibitor Cocktail
for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA
One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin
P2714-1BTL 1 bottle
for use in purification of Histidine-tagged proteins DMSO solution
Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon
One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml
P8849-1ML 1 mL
P8849-5ML 5 mL
Life Science Innovationsmdashnew and emerging technologies put forth in a fresh unique way that applies to your area of study
BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic
sigma-aldrichcom
Download your copy of Life Science Innovations at sigmacominnovations
Subscribe or download your copy of BioFiles at sigmacombiofiles
A Perfect Fit
Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you
Our Innovation Your Research mdash Shaping the Future of Life Science 7
Protease In
hib
itor C
ocktails an
d Tab
lets
for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators
One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver
P8340-1ML 1 mL
P8340-5ML 5 mL
for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators
Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins
Solution in DMSO (D 2650 Hybri-Max)
P1860-1ML 1 mL
for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA
Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli
Supplied with a vial of DMSO
P8465-5ML 5 mL
P8465-25ML 25 mL
for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 20 g of yeast
P8215-1ML 1 mL
P8215-5ML 5 mL
for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml
Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
P9599-1ML 1 mL
P9599-5ML 5 mL
While Sigma scientists have invested
considerable resources to formulate
the most effective cocktails suitable
for as many applications as possible
each cell line extractionpurification
procedure and expression system poses a different
set of proteolytic challenges We would like to solicit
feedback from researchers who have encountered
problems with standard cocktail formulations as
well as those who have developed novel alternative
strategies for protease inhibition Visit our BioBlog at
sigmabioblogscom to learn more
8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
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Your gateway to products services and more life science research
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Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
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of continually curated biological data
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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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Bro
ad S
pec
tru
m In
hib
ito
rs o
f Pr
ote
oly
tic
Enzy
me
Cla
sses
Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics
Typical Working Concentrations
Preparation of Stock Solutions
Serine Protease Inhibitors
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM 100 mM in water stable 1 month at -20 degC
A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC
D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC
T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl
T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Cysteine Protease Inhibitors
E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh
L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain
10-100 μM 10 mM in water stable 6 months at -20 degC
E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC
C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain
10-100 μM 10 mM in DMSO stable at -20 degC
A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain
1-100 μM 10 mM in water stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain
01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM
Aspartic Protease Inhibitors
P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
Metalloproteinase Inhibitors
ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water
P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC
R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase
1-10 mM 1 mM in water stable 1 month at -20 degC
B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC
M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC
References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data
Our Innovation Your Research mdash Shaping the Future of Life Science 5
Protease In
hib
itor Pan
el
Protease Inhibitor PanelProtease Inhibitor Panel
Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases
Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility
As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture
Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor
INHIB1-1KT 1 kit
INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility
Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location
Simple to use
Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)
capable of the cleavage
sigma-aldrichcom
Protease Finder
6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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ito
r C
ock
tails
an
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able
ts
Protease Inhibitor Cocktails and TabletsYou need a cocktail
End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg
For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)
One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract
S8820-20TAB 20 tablets
Protease Inhibitor Cocktail
for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA
One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin
P2714-1BTL 1 bottle
for use in purification of Histidine-tagged proteins DMSO solution
Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon
One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml
P8849-1ML 1 mL
P8849-5ML 5 mL
Life Science Innovationsmdashnew and emerging technologies put forth in a fresh unique way that applies to your area of study
BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic
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A Perfect Fit
Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you
Our Innovation Your Research mdash Shaping the Future of Life Science 7
Protease In
hib
itor C
ocktails an
d Tab
lets
for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators
One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver
P8340-1ML 1 mL
P8340-5ML 5 mL
for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators
Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins
Solution in DMSO (D 2650 Hybri-Max)
P1860-1ML 1 mL
for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA
Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli
Supplied with a vial of DMSO
P8465-5ML 5 mL
P8465-25ML 25 mL
for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 20 g of yeast
P8215-1ML 1 mL
P8215-5ML 5 mL
for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml
Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
P9599-1ML 1 mL
P9599-5ML 5 mL
While Sigma scientists have invested
considerable resources to formulate
the most effective cocktails suitable
for as many applications as possible
each cell line extractionpurification
procedure and expression system poses a different
set of proteolytic challenges We would like to solicit
feedback from researchers who have encountered
problems with standard cocktail formulations as
well as those who have developed novel alternative
strategies for protease inhibition Visit our BioBlog at
sigmabioblogscom to learn more
8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 5
Protease In
hib
itor Pan
el
Protease Inhibitor PanelProtease Inhibitor Panel
Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases
Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility
As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture
Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor
INHIB1-1KT 1 kit
INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility
Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location
Simple to use
Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)
capable of the cleavage
sigma-aldrichcom
Protease Finder
6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e In
hib
ito
r C
ock
tails
an
d T
able
ts
Protease Inhibitor Cocktails and TabletsYou need a cocktail
End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg
For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)
One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract
S8820-20TAB 20 tablets
Protease Inhibitor Cocktail
for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA
One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin
P2714-1BTL 1 bottle
for use in purification of Histidine-tagged proteins DMSO solution
Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon
One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml
P8849-1ML 1 mL
P8849-5ML 5 mL
Life Science Innovationsmdashnew and emerging technologies put forth in a fresh unique way that applies to your area of study
BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic
sigma-aldrichcom
Download your copy of Life Science Innovations at sigmacominnovations
Subscribe or download your copy of BioFiles at sigmacombiofiles
A Perfect Fit
Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you
Our Innovation Your Research mdash Shaping the Future of Life Science 7
Protease In
hib
itor C
ocktails an
d Tab
lets
for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators
One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver
P8340-1ML 1 mL
P8340-5ML 5 mL
for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators
Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins
Solution in DMSO (D 2650 Hybri-Max)
P1860-1ML 1 mL
for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA
Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli
Supplied with a vial of DMSO
P8465-5ML 5 mL
P8465-25ML 25 mL
for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 20 g of yeast
P8215-1ML 1 mL
P8215-5ML 5 mL
for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml
Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
P9599-1ML 1 mL
P9599-5ML 5 mL
While Sigma scientists have invested
considerable resources to formulate
the most effective cocktails suitable
for as many applications as possible
each cell line extractionpurification
procedure and expression system poses a different
set of proteolytic challenges We would like to solicit
feedback from researchers who have encountered
problems with standard cocktail formulations as
well as those who have developed novel alternative
strategies for protease inhibition Visit our BioBlog at
sigmabioblogscom to learn more
8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
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Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
Received your first research grantFocus on your discoveries ndash not your expenses Let Sigma-Aldrich help you make the most of your research dollars
Sigma-Aldrich New Lab Start-Up Program
bull Over 100000 quality biochemical and organic chemical products
bull Savings of up to 70
bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
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0039
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Pro
teas
e In
hib
ito
r C
ock
tails
an
d T
able
ts
Protease Inhibitor Cocktails and TabletsYou need a cocktail
End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg
For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)
One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract
S8820-20TAB 20 tablets
Protease Inhibitor Cocktail
for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA
One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin
P2714-1BTL 1 bottle
for use in purification of Histidine-tagged proteins DMSO solution
Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon
One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml
P8849-1ML 1 mL
P8849-5ML 5 mL
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Our Innovation Your Research mdash Shaping the Future of Life Science 7
Protease In
hib
itor C
ocktails an
d Tab
lets
for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators
One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver
P8340-1ML 1 mL
P8340-5ML 5 mL
for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators
Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins
Solution in DMSO (D 2650 Hybri-Max)
P1860-1ML 1 mL
for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA
Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli
Supplied with a vial of DMSO
P8465-5ML 5 mL
P8465-25ML 25 mL
for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 20 g of yeast
P8215-1ML 1 mL
P8215-5ML 5 mL
for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml
Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
P9599-1ML 1 mL
P9599-5ML 5 mL
While Sigma scientists have invested
considerable resources to formulate
the most effective cocktails suitable
for as many applications as possible
each cell line extractionpurification
procedure and expression system poses a different
set of proteolytic challenges We would like to solicit
feedback from researchers who have encountered
problems with standard cocktail formulations as
well as those who have developed novel alternative
strategies for protease inhibition Visit our BioBlog at
sigmabioblogscom to learn more
8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
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Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
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Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 7
Protease In
hib
itor C
ocktails an
d Tab
lets
for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators
One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver
P8340-1ML 1 mL
P8340-5ML 5 mL
for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators
Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins
Solution in DMSO (D 2650 Hybri-Max)
P1860-1ML 1 mL
for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA
Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli
Supplied with a vial of DMSO
P8465-5ML 5 mL
P8465-25ML 25 mL
for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 20 g of yeast
P8215-1ML 1 mL
P8215-5ML 5 mL
for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline
One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml
Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
No
Act
ivity
No
Act
ivity
12
1
08
06
04
02
0
Source
Aminopeptidase activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Am
ino
pep
tid
ase
acti
vity
(ug
tis
sue)
1
08
06
04
02
0
Source
Serine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
Seri
ne
pro
teas
e ac
tivi
ty(u
g t
issu
e)
03
025
02
015
01
005
0
Asp
arti
c p
rote
ase
acti
vity
(ug
tis
sue)
Source
Aspartic protease activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
00140012001
0008000600040002
0
Ch
ymo
tryp
sin
-lik
e ac
tivi
ty(u
g t
issu
e)
Source
Chymotrypsin-like activity
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
No
Act
ivity
218161412
108060402
0
Cys
tein
e p
rote
ase
acti
vity
(ug
tis
sue)
Source
Cysteine protease activity
as iswith inhibitor cocktail
beansprouts
peasprouts
pea leaves
pearoots
wheat sprouts
wheat leaves
wheat roots
tobacco sprouts
tobacco leaves
arabidopsis sprouts
Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail
P9599-1ML 1 mL
P9599-5ML 5 mL
While Sigma scientists have invested
considerable resources to formulate
the most effective cocktails suitable
for as many applications as possible
each cell line extractionpurification
procedure and expression system poses a different
set of proteolytic challenges We would like to solicit
feedback from researchers who have encountered
problems with standard cocktail formulations as
well as those who have developed novel alternative
strategies for protease inhibition Visit our BioBlog at
sigmabioblogscom to learn more
8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
Received your first research grantFocus on your discoveries ndash not your expenses Let Sigma-Aldrich help you make the most of your research dollars
Sigma-Aldrich New Lab Start-Up Program
bull Over 100000 quality biochemical and organic chemical products
bull Savings of up to 70
bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
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Sigma-Aldrich New Lab Start-Up Program
bull Over 100000 quality biochemical and organic chemical products
bull Savings of up to 70
bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
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powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
Received your first research grantFocus on your discoveries ndash not your expenses Let Sigma-Aldrich help you make the most of your research dollars
Sigma-Aldrich New Lab Start-Up Program
bull Over 100000 quality biochemical and organic chemical products
bull Savings of up to 70
bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
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of continually curated biological data
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
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N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
Received your first research grantFocus on your discoveries ndash not your expenses Let Sigma-Aldrich help you make the most of your research dollars
Sigma-Aldrich New Lab Start-Up Program
bull Over 100000 quality biochemical and organic chemical products
bull Savings of up to 70
bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
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N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
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bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)
ge95 powderA0835-1MG 1 mg
A0835-5MG 5 mg
Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42
ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide
A1341-1MG 1 mg
N-Acetyl-Ile-Glu-Thr-Asp-al
Ac-IETD-CHOC21H34N4O10 FW 50252
NH
HN
NH
HN
H
O
H3C
O
O
O
O
CH3
O OH
CH3HO
O
OH
CH3
~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3
A1216-1MG 1 mg
N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor
ge80 (HPLC) powderA1466-1MG 1 mg
Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes
N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases
Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM
ge97 (HPLC)A1276-250UG 250 μg
A1276-5MG 05 mg
A1276-1MG 1 mg
A1276-5MG 5 mg
A1276-10MG 10 mg
A1276-25MG 25 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
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bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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Our Innovation Your Research mdash Shaping the Future of Life Science 13
Prod
uct Listin
gs
2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus
Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5
Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)
ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator
EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature
4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride
AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO
H2N
F
bull HCl
Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM
Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin
Concentrations for 50 inhibition (μgml)
papain 016
trypsin 026
cathepsin A 119
cathepsin B 059
cathepsin D 125
plasmin gt93
chymotrypsin and pepsin gt250
It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM
Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC
Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization
Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC
A6191-1MG 1 mg
A6191-5MG 5 mg
A6191-25MG 25 mg
A6191-100MG 100 mg
14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
sigma-aldrichcom
Your gateway to products services and more life science research
Visit sigmacombiofiles
Read current and previous issues and register to receive future Biofiles issues
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
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Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
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14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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q Antithrombin III
Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase
Anti thrombin III from bovine plasma
lyophilized powder activity 200-400 unitsmg protein (E1
280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A9141-10UN 10 units
A9141-50UN 50 units
Anti thrombin III from human plasma
lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80
One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin
A2221-25UG 25 μg
A2221-125UG 125 μg
Anti thrombin III from rat plasma
lyophilized powder activity gt200 unitsmg protein (E1
280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75
One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin
Homogeneous by gel electrophoresis
A6397-10UN 10 units
Antithrombin III p
α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase
Chromatographically prepared and partially purified
salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A9024-5MG 5 mg
A9024-25MG 25 mg
A9024-100MG 100 mg
salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein
A6150-25MG 25 mg
A6150-100MG 100 mg
A6150-500MG 500 mg
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Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 15
Prod
uct Listin
gs
Aprotinin
10 20 30 40 50 60 70 80 90 100
Trypsin binding site
3 Disulfides
Signal Peptide
Aprotinin(bovine)
Propeptide
Propeptide
Kunitz domain
Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
Aprotinin Protease Inhibition Specificity
Enzyme Ki
Acrosin Weak inhibition
Chymotrypsin Ki = 9 nM
Chymotrypsinogen (bovine) pH 80 Ki = 9 nM
Elastase (human leukocytes) pH 80 Ki = 35 μM
Kallikrein (pancreatic) pH 80 Ki = 10 nM
Kallikrein (plasma) Ki = 30 nM 100 nM
Kallikrein (tissue) Ki = 1 nM
Kallikrein (urine) Ki = 17 nM
Plasmin (porcine) pH 78 Ki = 40 nM
Plasminogen activator Ki = 8 μM 27 μM
Trypsin (bovine) pH 80 Ki = 006 pM
Trypsinogen (bovine) pH 80 Ki = 18 μM
Tryptase TL-2 16 Inhibition at 10 μM
Urokinase (human) pH 88 Ki = 80 μM
q Aprotinin
Aprotinin bovine
BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)
Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods
65142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue
A6103-1MG 1 mg
A6103-25MG 25 mg
A6103-100MG 100 mg
Aprotinin (Bovine)
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
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Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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e D
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n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
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of continually curated biological data
and interactive genetic networks
all with a single simple click
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
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s
Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease
Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)
1 TIU sim 1300 KIU
saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol
BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter
A further purification of A1153 to yield a product with slightly higher activity and purity
One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC
65
142
20
2429
3645556697116
200
SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue
A6106-1MG 1 mg
A6106-25MG 25 mg
A6106-100MG 100 mg
Aprotinin p
Bdellin from leechesPlasmin inhibitor [62494-89-7]
lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B
Lyophilized powder containing glycine
Package size based on protein content
One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC
B3906-1MG 01 mg
Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money
Moving to a new locationMake a good first impression ndash choose high-quality products at competitive prices from Sigma-Aldrich
Received your first research grantFocus on your discoveries ndash not your expenses Let Sigma-Aldrich help you make the most of your research dollars
Sigma-Aldrich New Lab Start-Up Program
bull Over 100000 quality biochemical and organic chemical products
bull Savings of up to 70
bull Additional savings as your order value increases
Easy amp Economical
Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information
sigma-aldrichcom
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 17
Prod
uct Listin
gs
Benz ami dine hydrochloride hydrate
Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)
NH
NH2
bull HCl bull H2O
Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A
In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma
Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution
l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target
gt90 (TLC)B5306-1MG 1 mg
Bestatin hydrochloride
N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483
HN
O O
OH
H2NOH
HCl
CH3
CH3
A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin
ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC
B8385-5MG 05 mg
B8385-1MG 1 mg
B8385-5MG 5 mg
B8385-10MG 10 mg
B8385-25MG 25 mg
B8385-100MG 100 mg
Boc-Asp(OMe)-fluoro methyl keto ne
Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326
OCH3
ONH
OF
Boc
ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability
BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O
O
Br H
Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)
ge98B1552-5MG 5 mg
C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection
aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
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powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
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LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4
Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)
Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin
ge95 (HPLC)C9181-5MG 05 mg
C9181-1MG 1 mg
Captopril
N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N
O
OH
OHS
CH3
Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density
ge98 (HPLC) powderC4042-5G 5 g
C4042-25G 25 g
meets USP testing specificationsC8856-1G 1 g
C8856-5G 5 g
C8856-25G 25 g
Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]
lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC
Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts
Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde
A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile
chymostatin A MW = 6077
chymostatin B MW = 5937
chymostatin C MW = 6077
C7268-1MG 1 mg
C7268-5MG 5 mg
C7268-25MG 25 mg
C7268-50MG 50 mg
C7268-100MG 100 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 19
Prod
uct Listin
gs
clasto-Lactacystin β-lactone
[154226-60-5] C10H15NO4 FW 21323
NH
O
OH3C
H
OOH
CH3
CH3
Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone
L7035-1MG 01 mg
Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease
buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol
Protein determined by E1280
C0408-250UG 250 μg
C0408-5MG 05 mg
lyophilized powderPackage size based on protein content
Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases
D7910-5MG 5 mg
D7910-10MG 10 mg
D7910-25MG 25 mg
Dichloro methyl ene diphos pho nic acid disodium salt
Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa
O
OHP
Cl
ClHO
ONaO
Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14
Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)
D4434-1G 1 g
Diethyl enetri amine penta acetic acid
DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335
N
N
O
O
OO N
OH
OH
HO
HO
OH
O
ge99 (titration)Purified
D6518-5G 5 g
D6518-10G 10 g
D6518-50G 50 g
D6518-100G 100 g
Diiso pro pyl fluoro phos phate
DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415
CH3 CHCH3
O PO
FO CH
CH3
CH3
Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins
Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)
density 106 gmL 25 degC
D0879-1G 1 g
20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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s
α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
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LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity
E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)
Metallo-protease inhibitor with selectivity for aminopeptidases
ge97 (HPLC)E3389-1MG 1 mg
E3389-5MG 5 mg
q Ethylenediaminetetraacetic acid
Edathamil
Ethylene di amine tetra acetic acid disodium salt dihydrate
Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224
ORN
NRO
O
O
RO
O
OR
OR = 2 H and 2 Na
bull H2O
bull H2O
Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity
reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases
ED2SS-50G 50 g
ED2SS-100G 100 g
ED2SS-250G 250 g
ED2SS-500G 500 g
ED2SS-1KG 1 kg
ED2SS-25KG 25 kg
ED2SS-5KG 5 kg
Ethylene di amine tetra acetic acid tripotassium salt dihydrate
Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254
NN
KO
O
OK
OO
HO
OKO
bull 2H2O
Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals
98 (titration)E0270-25G 25 g
E0270-100G 100 g
Ethylenediaminetetraacetic acid p
Our Innovation Your Research mdash Shaping the Future of Life Science 21
A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
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of continually curated biological data
and interactive genetic networks
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
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A chelating agent useful for the determination of calcium in the presence of magnesium
ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level
E4378-10G 10 g
E4378-25G 25 g
E4378-100G 100 g
E4378-250G 250 g
E4378-500G 500 g
E4378-1KG 1 kg
N-Ethyl male imide
NEM [128-53-0] C6H7NO2 FW 12513 N
CH3
O O
Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels
Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases
Reagent for the covalent modification of cysteine residues in proteins
crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM
The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups
E3876-5G 5 g
E3876-25G 25 g
E3876-100G 100 g
NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70
Features over 600 detailed procedures for measuring enzyme activity and related metabolites
The Library is the result of over ten years of in-house work on analytical procedures developed
by Sigma-Aldrichreg scientists
22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
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to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
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0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase
gt93 (HPLC) solidG8291-5MG 05 mg
Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148
ge97 (HPLC)Useful for affinity chromatography
G5386-100MG 100 mg
Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration
Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC
One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
Protein determined by Lowry
lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)
H7016-10UN 10 units
H7016-100UN 100 units
H7016-500UN 500 units
H7016-2KU 2000 units
lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)
Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75
H7380-100UN 100 units
H7380-500UN 500 units
H7380-2KU 2000 units
[Lys47]Hirudin (HV2) leech[120300-65-4]
recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard
ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain
H6027-1MG 01 mg
Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines
ge97 (HPLC)I9759-5MG 5 mg
I9759-25MG 25 mg
Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt
Reported inhibitor of clostridial collagenase
gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg
Lactacystin
[133343-34-7] C15H24N2O7S FW 37643
NH
O
H3C HO
S OH
OH
H3C CH3
O
HN
O
O
CH3
ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
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du
ct L
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s
α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
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LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
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OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 23
Prod
uct Listin
gs
Leupeptin
N
O
O
N
O
N
O
H
H
H
H
CH2CH(CH3)2
CH2CH(CH3)2
CH2CH2CH2
CH3
CNH
NH
NH2
Leupeptin Protease Inhibitor Specificity
Enzyme Ki
Acrosin Ki = 210 nM
Calpains Ki = 10-430 nM
Cathepsin B Ki = 41 nM
Chymotrypsin Ki = 11 mM
Histolysin Ki = 43 nM
Plasmin Ki = 34 mM
Trypsin Ki = 35 nM
q Leupeptin
Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin
N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559
ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
Sigmalsquos commitment to life science research extends far beyond quality products
Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience
sigma-aldrichcom
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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e D
etec
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n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
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s
α2-Macro globulin from human plasma
α2-M
α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels
Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease
Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg
α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments
BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose
Package size based on protein content
One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein
65142
20
2429
36
45
5566
97116
200
Co
mp
etitor
Sigm
a
SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue
M6159-1MG 1 mg
M6159-5MG 5 mg
M6159-10MG 10 mg
N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne
N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299
Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources
Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide
Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing
A working concentration of 1 μM is stable for at least one day at room temperature
ge90 (HPLC)P5318-5MG 5 mg
P5318-25MG 25 mg
ge75 (HPLC)P4265-1MG 1 mg
P4265-5MG 5 mg
P4265-25MG 25 mg
P4265-100MG 100 mg
P4265-250MG 250 mg
Phenyl meth ane sul fonyl fluoride
PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419
S FO
O
ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75
P7626-250MG 250 mg
P7626-1G 1 g
P7626-5G 5 g
P7626-25G 25 g
P7626-100G 100 g
α2-Macroglobulin (Human Sequence)
Our Innovation Your Research mdash Shaping the Future of Life Science 25
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform
Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month
Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM
R7385-5MG 05 mg
R7385-1MG 1 mg
R7385-5MG 5 mg
R7385-10MG 10 mg
R7385-25MG 25 mg
Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor
ge95 (HPLC)P0350-5MG 05 mg
Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor
ge95 (HPLC)P0475-5MG 05 mg
gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6
ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli
T4376-100MG 100 mg
T4376-250MG 250 mg
T4376-1G 1 g
T4376-5G 5 g
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
du
ct L
isti
ng
s
Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo
There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive
This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary
Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent
Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]
10 20 30 40 50 60 70 80 90 100 110
Trypsin binding site
Chymotrypsin binding site
7 Disulfides
Signal Peptide
Propeptide
Trypsin Inhibitor (Bowman-Birk)(soybean)
Trypsin Inhibitor (Bowman-Birk) (soybean)
lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes
One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein
Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas
Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein
Protein determined by biuret
T0256-1MG 1 mg
T0256-5MG 5 mg
T0256-10MG 10 mg
T0256-25MG 25 mg
Trypsin inhibitor from chicken egg white
Ovo muco id
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
9 Disulfides
Signal Peptide
Trypsin Inhibitor (chicken egg white)
N-Linked Glycans
Kazal-like domains
Trypsin Inhibitor (chicken egg white)
Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein
T9253-250MG 250 mg
T9253-500MG 500 mg
T9253-1G 1 g
T9253-5G 5 g
Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein
T2011-250MG 250 mg
T2011-500MG 500 mg
T2011-1G 1 g
T2011-5G 5 g
Trypsin Inhibitor (Bowman-Birk) (soybean)
Trypsin Inhibitor (bovine pancreas)
Trypsin Inhibitor (chicken egg white)
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 27
Prod
uct Listin
gs
Trypsin inhibitor from Glycine max (soybean)
SBTI
Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108
Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color
20 40 60 80 100 120 140 160 180 200
Trypsin binding site
2 Disulfides
Signal Peptide
Trypsin Inhibitor (Kunitz)(soybean)
Propeptide
Trypsin Inhibitor (Kunitz) (soybean)
Type I-S lyophilized powderChromatographically prepared
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC
One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein
Protein determined by biuret
The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level
T9003-25MG 25 mg
T9003-100MG 100 mg
T9003-250MG 250 mg
T9003-500MG 500 mg
T9003-1G 1 g
T9003-5G 5 g
powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired
Type I-S
activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)
Chromatographically prepared
One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)
Protein determined by biuret
T6522-25MG 25 mg
T6522-100MG 100 mg
T6522-5X100MG 5 times 100 mg
T6522-250MG 250 mg
T6522-500MG 500 mg
T6522-1G 1 g
T6522-5G 5 g
lyophilized powderType II-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein
T9128-500MG 500 mg
T9128-1G 1 g
T9128-5G 5 g
T9128-10G 10 g
1 solution in water (contains no buffers)Prepared From Type I-S
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
Protein determined by biuret
T9008-5ML 5 mL
1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum
Optimized for passage of endothelial cell cultures
Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium
Endotoxin tested
T6414-100ML 100 mL
T6414-6X100ML 6 times 100 mL
Trypsin inhibitor from Phaseolus limensis (lima bean)
Trypsin Inhibitor from lima beans
Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization
Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)
One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path
One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein
T9378-100MG 100 mg
T9378-500MG 500 mg
T9378-1G 1 g
Trypsin inhibitor from turkey egg white
Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path
One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein
T4385-250MG 250 mg
T4385-1G 1 g
T4385-5G 5 g
Trypsin inhibitor p
Trypsin Inhibitor (Kunitz) (soybean)
28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex
ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc
powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition
C1730-1MG 1 mg
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Our Innovation Your Research mdash Shaping the Future of Life Science 29
Protease D
etection
Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer
Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity
The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays
Protease Fluorescent Detection Kit
The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate
Detection limit approx 5 ng trypsin
Supplied with sufficient reagents for up to 200 one mL assays
Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)
Add sample to microfuge tube
Add buffer and FITC-casein solution and incubate at 37degC for 60 min
Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate
Centrifuge
Remove Supernatant
Transfer dilute and read in a fluorimeter versus standard curve
010000
20000300004000050000600007000080000
0 5 10 15 20 25
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Nanograms of Trypsin
Typical Activity Curve Using Trypsin
0
10000
20000
30000
0 1 2 3 4 5 6 7
Nanograms of Trypsin
Flu
ore
scen
t U
nit
s (4
855
35 n
m)
Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin
Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys
Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions
Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo
Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro
Components
Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml
PF0100-1KT 1 kit
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience
Pro
teas
e D
etec
tio
n
Non-Specific Protease Substrates
Casein from bovine milk[9000-71-9]
technical gradeC7078-500G 500 g
C7078-1KG 1 kg
C7078-10KG 10 kg
Casein NN-dimethyl ated from bovine milk
essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein
Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)
Protease substrate
C9801-1G 1 g
C9801-5G 5 g
C9801-25G 25 g
C9801-50G 50 g
Casein fluor escein iso thio cyanate from bovine milk
FITC-casein
highly sensitive protease substrate
Type II essentially salt-free lyophilized powderC3777-25MG 25 mg
C3777-100MG 100 mg
Type III essentially salt-free lyophilized powderC0528-10MG 10 mg
C0528-25MG 25 mg
C0528-100MG 100 mg
Elastin-orcein
Elastin impregnated with orcein
Prepared from E1625
Substrate for the estimation of elastase
E1500-5G 5 g
ElastinndashCongo red
Elastin impregnated with Congo red dye
Substrate for the estimation of elastase
E0502-5G 5 g
Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry
~45 in H2OG7765-250ML 250 mL
G7765-1L 1 L
Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger
suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes
H2625-25G 25 g
H2625-100G 100 g
H2625-1KG 1 kg
HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R
Chromogenic substrate for trypsin
H6268-1G 1 g
H6268-5G 5 g
sigma-aldrichcom
Visit sigmacomenzymeexplorer
The New Enzyme Explorer
Redesigned homepage user interface
Expanded and reformatted technical resources and indexes
Protease Finder Identifies the cleavage sequence
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems
Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene
powered by Ingenuity gt sigmacomyfg
Imagine a tool that gives you the power
to navigate a comprehensive universe
of continually curated biological data
and interactive genetic networks
all with a single simple click
sigma-aldrichcom
LFL71112-508361
0039
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice
copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp
World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052
Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice