The TRA-1-85 monoclonal antibody (mAb) specifically binds to the high frequency blood group antigen OKa. The OKa blood group antigen is a 35-69 kDa glycoprotein also known as CD147, EMMPRIN, M6, or Basigin. The OKa group is widely expressed on most human tissues including, but not limited to kidney, peripheral blood leukocytes, liver, pancreas, colon, skin, and brain. TRA-1-85 mAb is often used to discern human cells from non-human cells in multiple applications including animal transplantation studies. During development, the purified TRA-1-85 mAb was found to detect OKa blood group antigen by western blot analysis of cellular lysates, by immunofluorescent staining of fixed cells, and by flow cytometric analysis of live cells. Flow cytometric analysis of TRA-1-85 expression on human cervical carcinoma (HeLa cell line). HeLa cells (ATCC® CCL 2™) were stained with Purified Mouse anti-Human TRA-1-85 Antigen monoclonal antibody (solid line histogram) or with Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121, dashed line histogram). The second-step reagent was FITC goat anti-Mouse Ig (Cat. No. 554001). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of the HeLa cell line. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System. Immunoflourescent staining of Tra-1-85 on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 47 grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were stained with Purified Mouse anti-Human TRA-1-85 Antigen monoclonal antibody (pseudo-colored green) at 1.2 μg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies), and cell nuclei were stained with Hoechst (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software.
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