TAF15 Antibody / TATA-binding protein-associated factor 2N (FY13059)

Western blot analysis of TAF15 using anti-TAF15 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human RT4 whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human whole cell lysates, Lane 4: human U251 whole cell lysates, Lane 5: rat testis tissue lysates, Lane 6: rat thymus tissue lysates, Lane 7: mouse testis tissue lysates, Lane 8: mouse thymus tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TAF15 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using an ECL Plus Western Blotting Substrate. A strong band is detected at approximately 68 kDa, slightly above the predicted molecular weight of 62 kDa. This apparent upward shift is consistent with published data showing that TAF15 commonly migrates at 66-70 kDa due to phosphorylation, arginine methylation, and its highly disordered N-terminal domain, all of which reduce SDS binding and slow gel mobility.