Next Generation Lectins Recombinant Lectins Ensure Stable Supplies Ready-to-Use Lectin Solution For Glycobiology & Cell Biology Wako has been developing lectins for sugar chain research, aiming to ensure those stable supplies. The first launch is recombinant BC2LCN (AiLecS1) for research of human induced pluripotent stem (iPS) and embryonic stem (ES) cells. rBC2LCN (AiLecS1) is expected to be applicable to not only stem cell research but also to regenerative medicine since it is reportedly a useful marker for detection of undifferentiated cells. rBC2LCN-FITC DAPI MERGE Apr. 2016 Application - Detections of an undifferentiation marker of human iPS cells Wako Pure Chemical Ind., Ltd. www.wako-chem.co.jp 1-2, Doshomachi 3-Chome Chuo-Ku, Osaka 540-8605, Japan Tel: 81-6-6203-3741 Fax: 81-6-6203-1999 Online Catalog: www.e-reagent.com Wako Chemicals USA, Inc. www.wakousa.com Toll-Free (U.S. only): 1-877-714-1920 Head Office (Richmond, VA): Tel: 1-804-714-1920/Fax: 1-804-271-7791 Los Angeles Sales Office (Irvine, CA): Tel: 1-949-679-1700/Fax: 1-949-679-1701 Boston Sales Office (Cambridge, MA): Tel: 1-617-354-6772/Fax: 1-617-354-6774 Wako Chemicals GmbH www.wako-chemicals.de Europe Office Fuggerstraße 12, D-41468 Neuss, Germany Tel: 49-2131-311-0 Fax: 49-2131-311100
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Next Generation Recombinant Lectins - Wako … Lectins Ensure Stable Supplies Ready-to-Use Lectin Solution For Glycobiology & Cell Biology Wako has been developing lectins for sugar
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Next Generation
Lectins
Recombinant Lectins Ensure Stable Supplies
Ready-to-Use Lectin Solution
For Glycobiology & Cell Biology
Wako has been developing lectins for sugar chain research, aiming to ensure those stable supplies. The first launch is recombinant BC2LCN (AiLecS1) for research of human induced pluripotent stem (iPS) and embryonic stem (ES) cells. rBC2LCN (AiLecS1) is expected to be applicable to not only stem cell research but also to regenerative medicine since it is reportedly a useful marker for detection of undifferentiated cells.
rBC2LCN-FITC DAPI MERGE
Apr. 2016
Application - Detections of an undifferentiation marker of human iPS cells
Wako Pure Chemical Ind., Ltd. www.wako-chem.co.jp
1-2, Doshomachi 3-Chome
Chuo-Ku, Osaka 540-8605, Japan
Tel: 81-6-6203-3741
Fax: 81-6-6203-1999
Online Catalog: www.e-reagent.com
Wako Chemicals USA, Inc. www.wakousa.com
Toll-Free (U.S. only): 1-877-714-1920
Head Office (Richmond, VA):
Tel: 1-804-714-1920/Fax: 1-804-271-7791
Los Angeles Sales Office (Irvine, CA):
Tel: 1-949-679-1700/Fax: 1-949-679-1701
Boston Sales Office (Cambridge, MA):
Tel: 1-617-354-6772/Fax: 1-617-354-6774
Wako Chemicals GmbH www.wako-chemicals.de
Europe Office
Fuggerstraße 12, D-41468
Neuss, Germany
Tel: 49-2131-311-0
Fax: 49-2131-311100
■ rBC2LCN (AiLecS1) from cDNA of Burkholderia cenocepacia
OCT3/4
TRA-1-81
rBC2LCN -Cy3
rBC2LCN -FITC
BSA-Cy3
BSA-FITC
DAPI MERGE
rBC2LCN 029-18061 (1mg), 025-18063 (1mg×5)
Glycan specificity Fucα1-2Galβ1-3GalNAc (H type 3) Fucα1-2Galβ1-3GlcNAc (H type 1)
Formulation PBS (pH 7.4)
cDNA source Synthetic cDNA based on Burkholderia cenocepacia
Protein concentration
1.2~1.8mg/ml (BCA Assay)
Source E. coli expressed Burkholderia cenocepacia lectin (BC2LCN/AiLecS1)
Molecular weight (theoretical value)
15,807
Purification Affinity purification by fucose agarose. Storage condition Keep at -20℃ before use. After thawing, keep at 2-10℃. Avoid the freeze-thaw cycle.
Product Outline
Detection of undifferentiated iPS cells using rBC2LCN
rBC2LCN References
rBC2LCN, a marker of undifferentiated human ES・iPS cells
Point in Check
rBC2LCN reportedly recognized an intramolecular sugar chain of Podocalyxin, exclusively, which exposes on the cell surface of undifferentiated human iPS cells. Please see the reference 1).
rBC2LCN Cy3
rBC2LCN FITC
DAPI MERGE BF
SSEA-4
TRA-1-81
<Detection condition> Fluorescent labeled rBC2LCN Final concentration : 0.1-0.5 μg/mL (1w/v% BSA, 1×PBS) Incubation time : 60 min. (RT, in the dark) Cell : Human iPS 201B7 strain Fixing condition: 4w/v% Paraformaldehyde for 10min. at RT or 4℃.
Detection of undifferentiated marker in human iPS cell colony naturally differentiated
Point in Check rBC2LCN (rLectin)
Anti TRA-1-81 Mab (Mouse IgM)
Anti SSEA-4 Mab (Mouse IgG3)
M. W. (kDa) 16 900 150
Flow Cytometry ○ ○ ○
Cytological staining ○ ○ ○
Live cell imaging ○ △ - Non-specific binding (Cytological staining)
Low High Low
1) Tateno, H., Matsushima, A., Hiemori, K., Onuma, Y., Ito, Y., Hasehira, K., Nishimura, K., Ohtaka, M., Takayasu, S., Nakanishi, M., Ikehara, Y., Nakanishi, M., Ohnuma, K., Chan, T., Toyoda, M., Akatsu, H., Umezawa, A., Asashima, M. and Hirabayashi, J. “Podocalyxin is a Glycoprotein ligand of the human pluripotent stem cell-specific probe rBC2LCN.”, Stem Cells Translational Medicine, 2(4), 265-73(2013). 2) Onuma, Y., Tateno, H., Hirabayashi, J., Ito, Y. and Asashima, M. “rBC2LCN, a new probe for live cell imaging of human pluripotent stem cells.”, Biochem. Biophys. Res. Commun., 431(3), 524-9(2013). 3) Tateno, H., Toyota, M., Saito, S., Onuma, Y., Ito, Y., Hiemori, K., Fukumura, M., Matsushima, A., Nakanishi, M., Ohnuma, K., Akutsu, H., Umezawa, A., Horimoto, K., Hirabayashi, J. and Asashima, M. “Glycome diagnosis of human induced pluripotent stem cells using lectin microarray.”, J. Biol. Chem., 286(23), 20345-53(2011). 4) Tateno, H., Onuma, Y., Ito, Y., Hiemori, K., Aiki, Y., Shimizu, M., Higuchi, K., Fukuda, M., Warashina, M., Honda, S., Asashima, M. and Hirabayashi, J. “A medium hyperglycosylated podocalyxin enables noninvasiva and quantitative detection of tumorigenic human pluripotent stem cells.”, Sci. Rep., 4, 4069(2014).
*1: Lectin group means one kind of classification of expediency based on the glycan specificity. It doesn’t warrant the recognition ability of all glycans.
*2: The information of the glycan specificity are the common information. These don’t limit the specificity of lectins.
Lectin Groups*1 Name Code No. Package
Size
cDNA Source
Glycan Specificity *2
Fucose-
Complex
BC2LCN
(AiLecS1)
029-18061
025-18063 1mg
1mg×5
cDNA Source : Burkholderia cenocepacia
Target Glycan : Fucα1-2Galβ1-3GalNAc (H type 3), Fucα1-2Galβ1-3GlcNAc (H type 1)