Stemgent® StainAlive™ DyLight™ 488 Mouse anti-Human TRA-1-60 Antibody

The Stemgent® StainAlive™ DyLight™ 488 Mouse anti-Human TRA-1-60 Antibody was screened on live, unfixed human embryonic stem (ES) cells using immunocytochemistry (ICC). This antibody was selected as the best conjugated TRA-1-60 antibody available for visualization of ES/iPS (induced pluripotent stem) cells, utilizing direct fluorescent staining and real-time imaging without fixation or killing the cells. This antibody format is useful for direct visualization of the reprogramming process to iPS colonies. For use on live cells, this antibody has been specifically formulated with low endotoxin and no sodium azide and is confirmed to be free of bacteria, fungi, mouse virus and mycoplasma as tested. The live ES cell morphology, proliferation and expression of pluripotency markers did not change after staining with this antibody. Clone TRA-1-60 reacts with a pluripotent-stem-cell-specific antigen expressed on undifferentiated human embryonic stem (ES) cells, embryonal carcinoma (EC) cells, and embryonic germ (EG) cells. The expression of TRA-1-60 on human ES cells is down-regulated upon differentiation. The TRA-1-60 antibody recognizes a neuraminidase-resistant carbohydrate epitope expressed on podocalyxin, a member of the CD34-related family of sialomucins. Podocalyxin is a transmembrane glycoprotein, which has been implicated in the development of aggressiveness in a variety of cancers, including breast and prostate cancer.

1. Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., and Yamanaka, S. (2007). Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors. Cell 131: 861-872.
2. Yu J., Vodyanik M.A., Smuga-Otto K., Antosiewicz-Bourget J., Frane J.L., Tian S., Nie J., Jonsdottir G.A., Ruotti V., Stewart R., Slukvin I.I., Thomson J.A. (2007). Induced pluripotent stem cell lines derived from human somatic cells. Science 318: 1917-1920.
3. Schopperle W.M., DeWolf W.C. (2007). The TRA-1-60 and TRA-1-81 human pluripotent stem cell markers are expressed on podocalyxin in embryonal carcinoma. Stem Cells 25: 723-730.
4. Chin A.C., Fong W.J., Goh L.T., Philp R., Oh S.K., Choo A.B. (2007) Identification of proteins from feeder conditioned medium that support human embryonic stem cells. J Biotechnol. 130:320-328.
5. Badcock G., Pigott C., Goepel J., Andrews P.W. (1999). The human embryonal carcinoma marker antigen TRA-1-60 is a sialylated keratan sulfate proteoglycan. Cancer Res. 59: 4715-4719.
6. Andrews PW, Banting G, Damjanov I, Arnaud D, Avner P. (1984). Three monoclonal antibodies defining distinct differentiation antigens associated with different high molecular weight polypeptides on the surface of human embryonal carcinoma cells. Hybridoma 3:347.
7. Lowry, W.E., Richter, L., Yachechko, R., Pyle, A.D., Tchieu, J., Sridharan, R., Clark, A.T., Plath, K. (2008) Generation of human induced pluripotent stem cells from dermal fibroblasts. Proc Natl Acad Sci U S A 105: 2883-8.