Immunofluorescent analysis of TRA-1-81 using anti-TRA-1-81 monoclonal antibody (Product # MA1-024) shows staining on the cell surface of human HEL 11.4 iPS cells, indicating pluripotency. TRA-1-81 staining (green) and an overlay image of TRA-1-81 with DAPI (blue) is shown. HEL 11.4 iPS cells were grown on matrigel coated chamber slides and fixed with formaldehyde prior to staining. Cells were probed with a Monoclonal antibody recognizing TRA-1-81 (Product # MA1-024) at a dilution of 1:20 overnight at 4 ºC, washed with PBS and incubated with a fluorescein conjugated secondary antibody at a dilution of 1:100 for 60 minutes at room temperature. Images were taken at 20X magnification.

Western blot analysis of TRA-1-81 was performed by loading 100ug of NTERA-2 whole cell lysate onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. Membranes were probed with a mouse monoclonal antibody recognizing TRA-1-81 (Product # MA1-024) at a dilution of 1:1500 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-IgM HRP secondary antibody at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Super Signal West Dura (Product #34075).