www.takara.co.kr Clontech 판매 주년 기념 tdTomato GFP Figure 1. Subcellular labeling. AcGFP1 and DsRed2 protein fusions were transiently trans- fected and visualized by fluorescence microscopy. Panel A. pAcGFP1-Mito (mitochondria) and pDsRed2-Nuc (nucleus) in HEK 293 cells. Panel B. pAcGFP1-Golgi (Golgi apparatus) and pDsRed2-Nuc (nucleus) in HEK 293 cells. Figure 2. In vivo Imaging. False-color overlay images (regions of interest encircled) highlight tdTomato fluorescence, but not GFP fluorescence, in the cadaver model. Panel A. Implanted tube with 100 x 10 6 MDA-MB-231-tdTomato-expressing cells, imaged with the DsRed filter set. Panel B. Implanted tube with 100 x 10 6 MDA-MB-231-GFP-expressing cells, imaged with the GFP filter set. Exposure times: 1 sec. A B A B C L O N T E C H C A M P A I G N E • Full color spectrum-cyan부터 far red까지 총 26 종 보유 • 최적화된 형광 & photostability • 다양한 application 가능 • EGFP보다 뛰어난 성능의 AcGFP 보유 • Multicolor imaging에 적합 • Live cell에서 쉽게 검출 가능 • Specific한 fluorescent protein antibody 보유 Application에 따른 선택 가이드 Application 추천 제품 비고 Protein localization • N- 또는 C- 말단에 fusion AcGFP1, DsRed-Monomer, mCherry 단백질은 monomeric 형태로 • 다양한 형광 단백질 (cyan to far red) 타겟 단백질의 기능을 방해를 줄일 수 있어서 fusion 단백질 실험에 이상적 Bacterial expression • 다양한 bacterial expression vector 보유 Lac promoter 이용 Subcellular labeling • AcGFP1 세포 소기관을 표식할 수 있는 다양한 형태의 vector 보유 • DsRed-Monomer (golgi complexes,actin filaments, plasma membranes, nuclei, • DsRed2 microtubules, endoplasmic reticuli, mitochondria, Peroxisomes, • mCherry endosomes) (Figure 1). • HcRed1 In vivo imaging • mPlum • E2-Crimson In vivo imaging에는 백그라운드를 줄여줄 수 있는 red나 far red • HcRed 형광 단백질 추천 (Figure 2). • tdTomato Fluorescence • AcGFP1 & DsRed-Monomer resonance energy • AcGFP1 & mCherry FRET은 두 단백질의 상호작용을 모니터 가능 transfer • mOrange & mStrawberry 성공적인 FRET실험을 위해서는 올바른 pair 선택이 중요 (FRET) • mOrange & mCherry Protein dynamics & • PAmCherry - 350-400nm의 빛을 Photoactivatable과 photoconvertible 형광 단백질은 half-life와 half & life 조사하면 형광을 띰 trafficking을 연구할 수 있는 강력한 방법 • Dendra2 - UV 또는 푸른 빛을 조사하면 녹색에서 붉은색으로 변화 파격 소비자 할인가 165,000원 * 제품 리스트 참조 (VAT 별도) Vector 한정 수량 Living Colors Fluorescent Proteins 다수의 citation을 보유하고 있는
6
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042-828-6525
www.takara.co.kr
Clontech 판매 주년 기념
3
F L U O R E S C E N T P R O T E I N S & R E P O R T E R S
tdTomato GFP
Applications of Living Colors Fluorescent ProteinsClontech’s fluorescent proteins are available in plasmid, lentiviral, and retroviral vector formats for a range of key applications. Visit www.clontech.com for a complete list of our vectors.
Fusions
FRET
Cellular & subcellular labeling
Gene expression
Kinetic studies
Cell capture
Cell cycle reporters
Proteasome activity analysis
Promoter reporters
Timed gene expression
In vivo imaging
Stem cell applications
Figure 1. Subcellular labeling. AcGFP1 and DsRed2 protein fusions were transiently trans-fected and visualized by fluorescence microscopy. Panel A. pAcGFP1-Mito (mitochondria) and pDsRed2-Nuc (nucleus) in HEK 293 cells. Panel B. pAcGFP1-Golgi (Golgi apparatus) and pDsRed2-Nuc (nucleus) in HEK 293 cells.
Figure 2. In vivo Imaging. False-color overlay images (regions of interest encircled) highlight tdTomato fluorescence, but not GFP fluorescence, in the cadaver model. Panel A. Implanted tube with 100 x 106 MDA-MB-231-tdTomato-expressing cells, imaged with the DsRed filter set. Panel B. Implanted tube with 100 x 106 MDA-MB-231-GFP-expressing cells, imaged with the GFP filter set. Exposure times: 1 sec.
A B A B
Selected Applications of Living Colors Fluorescent Proteins
Application Recommended Products Comments
Protein localization Fusion fluorescent proteins from far red to cyan are available in a variety of vector formats for both N- and C-terminal fusions.
AcGFP1, DsRed-Monomer, and mCherry proteins are ideal for fusions, since they are monomeric and least disruptive to the function of the protein of interest.
Bacterial expression We offer a variety of basic fluorescent protein vectors for bacterial expression.
The bacterial expression of fluorescent proteins is driven by the lac promoter.
FRET can be used to monitor the proximity and interaction of two proteins. Success of FRET relies on the correct pairing of donor and acceptor fluorescent proteins.
Protein dynamics & half-life PAmCherry—fluoresces when activated by 350–400 nm light.
Dendra2—converts from green to red when exposed to UV or blue light.
Photoactivatable and photoconvertible fluorescent proteins are powerful tools for studying protein half-life and trafficking, because they only mark molecules that were present at the time of activation.
trafficking을 연구할 수 있는 강력한 방법 • Dendra2 - UV 또는 푸른 빛을 조사하면
녹색에서 붉은색으로 변화
파격 소비자 할인가 165,000원
* 제품 리스트 참조 (VAT 별도)
Vector 한정
수량
Living Colors Fluorescent Proteins다수의 citation을 보유하고 있는
Clontech vector(FP)에는 Clontech 항체가 최적
Clontech에서는 다양한 fluorescent vector와 함께 specific한 antibody를 보유하고 있습니다. Clontech의 항체는 western blot, immunoprecipitation, immu-
nocytochemistry를 포함하여 다양한 실험에 적용 가능합니다.
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F L U O R E S C E N T P R O T E I N S & R E P O R T E R S
AntibodiesClontech offers highly specific antibodies for the detection of our Living Colors fluorescent proteins. Our antibodies are compatible with many applications, including Western blotting, immunoprecipitation, and immunocytochemistry.
EGFP Monoclonal
Antibody
Full-Length ZsGreen
Polyclonal Antibody
A.v. Monoclonal
Antibody (JL-8)
Full-Length GFP
Polyclonal Antibody
A.v. Peptide Antibody
GFP Monoclonal
Antibody
Proven Usesa WB, IP, IC WB WB, IP, IC WB WB, IP, IC WB, IP, IC
AcGFP1
ZsGreen1
GFP
GFPuv
EGFPb
EYFPb
ECFPb
a WB = Western Blot, IP = immunoprecipitation, IC = immunocytochemistry
b Not sold by Clontech
Antibody Selection Guide for Living Colors Fluorescent Proteins
termination sequence)와 WPRE (woodchuck hepatitis virus post transcriptional
regulatory element)를 포함하여 고역가의 렌티바이러스 조제 가능하며, 타겟 세포
로의 transduction과 유전자 발현을 증가시킨다.
* 다양한 형태의 lentiviral vector 보유, 뒷장 제품 리스트 참조
최적화된 Lenti-X™ Vector와 발현 시스템
03 / 04
5' LTR WPRE
RREcPPT/CTS
Core lentiviral vector backbone
Vector-specific elements
3' LTR
Lentiviral vector의 핵심 부분
O P T I M I Z E D L E N T I - X V E C T O R S Y S T E M S F O R A N Y A P P L I C A T I O N
34
PurorPEF1
PEF1 FP
FP = fluorescent protein (mCherry or ZsGreen1)
MC
S
pLVX-EF1 -IRES-Puro*
pLVX-EF1 -IRES-FP**
PEF1 FP
PEF1 FP
FP = fluorescent protein (AcGFP1, DsRed-Monomer, or mCherry)
MC
S
MC
S
pLVX-EF1 -FP-N1**
pLVX-EF1 -FP-C1**
IRES
MC
S
IRES
PPGKPPGK
PPGK Puror
Puror
Neo/HygPCMV IE IRES Puror
FP = fluorescent protein (mCherry, tdTomato, or ZsGreen1)
PCMV IE IRES FP
MC
SM
CS
pLVX-IRES-Puro/Neo/Hyg*
pLVX-IRES-FP*
Constitutive cDNA Expression
PCMV IE PurorPPGKMC
S
pLVX-Puro*
shRNA Expression
PurorPU6 PPGK
PU6 PCMV IE ZsGreen1
MC
SM
CS
pLVX-shRNA1*
pLVX-shRNA2**
Tetracycline-Inducible Expression
ProteoTuner™ Protein Control
Bicistronic Expression
Fluorescent Protein Expression
EF-1 Alpha Promoter
Promoter Reporters
PCMV IE DD IRES Puror
PCMV IE IRES ZsGreen1DD
MC
SM
CS
pLVX-PTuner**
pLVX-PTuner Green**
DD
FP = fluorescent protein (AmCyan1, ZsGreen1, or tdTomato)
MetLuc
FP
MC
SM
CS
pLVX-MetLuc*
pLVX-DD-FP*
PCMV IE PurorPPGKFP
PCMV IE PurorPPGKFP
FP = fluorescent protein (AcGFP1, AmCyan1, DsRed-Express2,DsRed-Monomer, mCherry, tdTomato, or ZsGreen1)
MC
S
MC
S
pLVX-FP-N1**
pLVX-FP-C1**
pLVX-Tet3G
pLVX-TRE3G
PCMV IE Tet-On 3G IRES Neor
PurorPTRE3GV PPGKMC
S
DD FP
MC
S
pLVX-DD-FP*
형광단백질이나 분비형 luciferase를 발현시키는 리포터시스템을 사용하여 promoter의 활성을 모니터 할 수 있다.
ProteoTuner destabilization domain(DD) 뒤쪽에 목적 유전자를 클로닝하면 리간드ligand인 Shield1을 이용하여 목적유전자의 안정화를 조절할 수 있다. Shield1은 소분자 리간드로 DD와 융합된 단백질이 proteosomal degradation되는 것을 방지하여 “protein on”으로 세포에 축적되도록 한다.
U6 promoter 뒤쪽으로 목적 shRNA 서열을 클로닝하면 트랜스펙션 시키기 어려운 세포에서도 유전자 발현을 knockdown시킬 수 있다.
Tet-On 3G나 Tet-Express vector system을 사용하면 정밀하고 높은 유전자 유도발현을 가능하게 한다.CMV promoter를 이용해 목적유전자를 발현시키고 항생제 마커를 활용해 렌티바이러스가 integration된 세포를 선별한다.
IRES가 포함된 렌티바이러스 vector를 이용하면 하나의 전사산물에서 목적유전자와 또 하나의 유전자(항생제 내성 유전자 또는 형광발현 유전자)를 공발현시킬 수 있다.
다양한 형광단백질(FP) 중 하나가 MCS의 N-말단 또는 C-말단에 클로닝 되어있어 목적 유전자가 단백질로 발현되고 세포내 위치하는 것을 모니터 할 수 있게 한다.
* Vector는 system 구성품으로만 판매된다.** Vector는 별도 구매 가능하다.
EF-1 α promoter를 이용하면 조혈모세포나 줄기세포와 같은 세포에서 CMV promoter 사용시 나타나는 silencing을 극복하고 목적유전자를 발현시킬 수 있다.
FP = fluorescent protein (AmCyan1, ZsGreen1, or tdTomato)
Promoter 특징
•CMV : 도입한 유전자를 항상 안정하게 발현한다
•EF-1α : 다른 promoter(CMV 등)로 발현이 낮거나, 줄기세포와 같이 promoter가 silencing이 되는 경우 유용하다.
※ PGK promoter의 하류에 항생제 내성 유전자(neo, hyg, puro 내성)가 포함되어 있어 stable cell line 선별에 이용할 수 있다.
형광단백질의 특징
•AcGFP1(monomer) : 목적단백질과의 융합 발현에 적합하다.
•ZsGreen1(Tetramer) : 유전자 도입효율을 모니터하는 리포터 발현에 적합하다. (IRES서열에 의해 목적유전자와 별도로 각각 번역된다)
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최고의 Lentivirus delivery를 위한, 최적화된 Lentiviral Vector 한정
수량
T e t r a c y c l i n e I n d u c i b l e E x p r e s s i o n
•귀중한 세포를 살아있는 상태로 실험에 이용 _ Transfection이 어렵거나 잘 성장하지 않는 세포도 살아있는 상태로 배양하면서 시간의 흐름에 따른 데이터 획득이 가능
•비용 절감 _ 동일세포로반복실험이가능하기때문에추가적인세포배양/세포용해등의과정이불필요
Cell lysis 과정
없이 정확하고
안정적인 실험!
The FB14 represents a unique portable high performance luminometer.
ConceptThe FB14 is a very small, portable single tube luminometer that combines excellent performance with ease of use and versatility. Its built-in software is ideal for field measurements. When used in the lab, a powerful PC software can be added for numerous data analysis functions.
Detection SystemThe sample compartment is situated inside a sliding door, which initiates the measurement sequence upon closing. The proximity of the sample to the light detector and the use of a light reflector surrounding the major part of the vial produce optimal photon collection.The resulting efficiency, combined with the high-speed photon counter, provide a measurement performance which is unique in its class.
User Interface OptionsThe FB14 can be run from its embedded software, but can also be interfaced with a powerful PC-software package:The Single Tube Luminometer PC Software is designed for stationary use in the laboratory. In addition to the Quick Measurement function, it offers five additional protocol types: Single Assay, Dual Assay, Single Kinetics, Multiple Kinetics and Cut-off Assay. The operator can define new protocols of all five types. Highly flexible graphical presentation of kinetic data is supported.
Advanced Tools for The Life Sciences www.titertek-berthold.com
FB14 Portable Tube Luminometer
The FB14 can be used for any luminescent glow-type
application performed in single tube format, for example: · Water Testing
· ATP Assays
· Immunoassays
· DNA / RNA Probe Assays
· Toxicity Assays
· Biothreat Assays
ConceptThe FB12 system is ideal for measurement of glow type luminescence fromsingle samples. Due to the unique design, a variety of sample formats can be accommodated. FB12 Luminometer combines a very small footprintwith excellent sensitivity and outstanding ease of use.In stand-alone mode, FB12 is ready for standard measurements. Samplesare readily loaded into the retractable drawer. Closing the sample drawerautomatically initiates the measurement. An external printer may be connectedoptionally.The modular Windows® Luminometer PC-Software or the Pocket PC-Lumino-meter Software can further enhance performance by organizing the measure-ment dialog and data evaluation.
ApplicationsFB12 is the instrument of choice for all luminescent glow type applicationsperformed in single sample format, for example:· Reporter gene assays, including dual-reporter assays· ATP assays · Cell proliferation, cytotoxicity and biomass assays, apoptosis assays· Detection of mycoplasma contamination in cell culture· Cellular luminescence/reactive oxygen species· Environmental toxicity and mutagenicity assays
DetectorThe FB12 Luminometer features advanced photoncounting electronics coupledwith a highly sensitive detector, providing the lowest signal backgroundand highest linearity spanning 6 decades. No cumbersome internal calibrationis required and no gain switch is needed.Light collection is maximized through both the proximity of the sample to thedetector and the use of optimized light reflectors surrounding the samplevial. As a result, reporter gene assays using firefly luciferase can now detectless than 1000 molecules of the enzyme with the FB12 Luminometer.
Stand-alone Operation FB12 offers basic raw data, extended raw data and Cut-off measurementprotocols. The interface supports the addition of an external serial printer.Three softkeys allow easy and intuitive operation. Both delay and measure-ment time can be set in steps of 0.1s.
Windows PC Software PlatformThe capabilities of the FB12 Luminometer are enhanced by its PC-Software.Even complex tasks can be performed conveniently using the PC program whichruns on a Microsoft® Windows® platform. It provides a direct link to the Micro-soft® Excel spreadsheet program. All suitable Windows® features are supported.The software relies on open system architecture:The basic module for standard measurements meets most basic needs.For more demanding tasks, Single Assay, Dual Assay, Single Kinetics, MultipleKinetics and Cut-off Assay protocol types are available.
FB12 Performance and Ease of Use
Versatile sample formats
The Sirius L is a compact high performance tube luminometer with unique features:
The sample versatility is unmatched.
The Quickstart capabilities provide results on the fly.
The intuitive and powerful PC software sets the standard:
Windows®7 compatible, self-detecting USB-port.
Sirius L in Flash configuration (left) and Glow configuration (right)
Wide Range of ApplicationsThe Sirius L is the perfect choice for all luminescent flash and glow applications performed in single sample format, for example:
· Reporter gene assays, including dual-reporter assays· Rapid detection of mycoplasma contamination in cell cultures: Lonza MycoAlert®
· ATP assays· Cell proliferation, cytotoxicity and biomass assays· Apoptosis assays· Cellular luminescence/reactive oxygen species· Environmental toxicity and mutagenicity assays
Supersensitive DetectorThe Sirius L detects less than 0.5 zmol of firefly luciferase. Light collection is maximized through both the proximity of the sample to the detector and the use of optimized light reflectors surrounding the sample vial.
Quickstart CapabilitiesClosing of the sample drawer automatically initiates the measurement with no need to press any key. The automatic light shutter protects the detector from ambient light, making the instrument ready to measure from the moment the door is closed.
Sample FormatsThe sample chamber is designed for performance and flexibility. Accomodation of a variety of vial types: From 35 mm petri dishes to micro-fuge format to 20 ml vials, there is virtually no limit.
Various Injector OptionsThe instrument is available with up to two syringe-type automatic reagent injectors. The injection volume ranges from 20–500 µl. Ready-to go sample holders for precise injection are available for 1.5 ml and 2.5 ml microfuge tubes, as well as for 55 mm and 75 mm, 12 mm diameter tubes (standard format for most luminescence assays).