Anti Keratan Sulfate (KS/Keratosulfate) lacking oversulfated structures mAb (Clone R-10G)

Cosmo Bio

SKU:
CAC-RIT-M001
  • Table. Comparison of R-10G antibody and existing iPS/ES marker antibody. iPS cells, induced pluripotent stem cells; ES cells, embryonic stem cells; EC cells, embryonal carcinoma cells. TRA-1-60, TRA-1-81 are from: Schopperle Et Al , WM , Stem Cells 2007 Mar; 25 (3): 723-30.
  • Figure 1. Pluripotent cell epitopes on the Podocalyxin molecular structure.
  • Figure 2. Screening of R-10G mAb by western blotting. Tic cell lysates in complete RIPA buffer were resolved by SDS-PAGE on a 4-15% gradient gel under nonreducing conditions, followed by immunoblot detection with R-10G. A) GelCode* Blue staining of SDS-PAGE gel containing Tic cell lysates (10ug protein). B) Tic cell lysates were analyzed by western blotting with R-10G. Molecular mass markers are shown on the left.
  • Figure 3. Localization of R-10G and TRA-1-81 epitopes on cultured Tic (hiPS) cells visualized by laser confocal microscopy. Tic cells cultured on Millicell® EZ slides were double-stained with R-10G and Alexa Fluor® 488-conjugated secondary (anti-mouse IgG1) antibody, followed by TRA-1-81 and Alexa Fluor® 555-conjugated secondary (anti-mouse IgM) antibody. Cells were observed at two different magnifications: ×100 (upper panel) and ×400 (lower panel). (I) Nomarski imaging. (II) Nuclear counterstaining with TO-PRO®-3. (III) Antigens for R-10G (green). (IV) Antigens for TRA-1-81 (conventional hiPS marker antibody) (red). (V) Merged image of (III) and (IV). (VI) Close-up view of V (×400). GelCode® is a registered trademarks of Thermo Fisher Scientific Co. and the subsidiary company's trademark. Millicell® is a registered trademarks of Merck Millipore (Merck KGaA). Alexa Fluor® and TO-PRO®a registered trademarks of Life Technologies Corporation. Ritsumeikan University Glycoengineering Research Center Toshiya Kawashima
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Monoclonal antibody R-10G recognizes keratan sulfate lacking oversulfated structures in oligosaccharide segments of keratan sulfate glycosaminoglycan chains on human induced pluripotent stem (hiPS) cells. Digestion with Keratanase II, Keratanase or Endo-beta-galactosidase removes these epitopes from keratan sulfate proteoglycans. Unlike commercially available keratan sulfate-recognizing antibodies 5D4 and BCD4, R-10G recognizes keratan sulfate with a low degree of sulfation. The antigenic polypeptide portion has been identified as podocalyxin. Importantly, R-10G antibody binds to human iPS cells and human ES cells, but only at very low levels to human embryonal carcinoma (EC) cells. Unlike widely used human pluripotent stem cell marker antibodies TRA-1-60 (IgM) and TRA-1-81 (IgM), R-10G (IgG1) binds only weakly to EC cells. Therefore, R-10G is useful for quality control of human iPS and ES cells.

References:
1) Kawabe K, et al, A novel antibody for human-induced pluripotent stem cells and embryonic stem cells recognizes a type of keratan sulfate lacking oversulfated structures. Glycobiology. 2013 Mar;23(3):322-36. PubMed: 23154990
2) Schopperle WM, et al, The TRA-1-60 and TRA-1-81 human pluripotent stem cell markers are expressed on podocalyxin in embryonal carcinoma. Stem Cells. 2007 Mar;25(3):723-30. PMID: 17124010



Product Specifications
Application IF, WB
Reactivity Human
Clonality Monoclonal (Clone No.: R-10G)
Host Mouse


Documents & Links for Anti Keratan Sulfate (KS/Keratosulfate) lacking oversulfated structures mAb (Clone R-10G)
Datasheet Anti Keratan Sulfate (KS/Keratosulfate) mAb (Clone R-10G) Datasheet

Documents & Links for Anti Keratan Sulfate (KS/Keratosulfate) lacking oversulfated structures mAb (Clone R-10G)
Datasheet Anti Keratan Sulfate (KS/Keratosulfate) mAb (Clone R-10G) Datasheet



Citations for Anti Keratan Sulfate (KS/Keratosulfate) lacking oversulfated structures mAb (Clone R-10G) – 2 Found
Mawaribuchi, Shuuji; Aiki, Yasuhiko; Ikeda, Nozomi; Ito, Yuzuru. mRNA and miRNA expression profiles in an ectoderm-biased substate of human pluripotent stem cells. Scientific Reports. 2019;9(1):11910.  PubMed
- Comments: Initial description of R-10G monoclonal antibody
Kawabe, Keiko; Tateyama, Daiki; Toyoda, Hidenao; Kawasaki, Nana; Hashii, Noritaka; Nakao, Hiromi; Matsumoto, Shogo; Nonaka, Motohiro; Matsumura, Hiroko; Hirose, Yoshinori; Morita, Ayaha; Katayama, Madoka; Sakuma, Makoto; Kawasaki, Nobuko; Furue, Miho Kusuda; Kawasaki, Toshisuke. A novel antibody for human induced pluripotent stem cells and embryonic stem cells recognizes a type of keratan sulfate lacking oversulfated structures. Glycobiology. 2013;23(3):322-36.  PubMed
- Comments: Initial description of R-10G monoclonal antibody
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