SETX Antibody / Senataxin (FY12819)

Western blot analysis of SETX using anti-SETX antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: rat L6 whole cell lysates, Lane 2: mouse C2C12 whole cell 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-SETX 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 band is observed near ~300 kDa together with higher-molecular-weight species forming a ladder. These upper bands are consistent with poly-ubiquitinated/SUMO-modified or complexed forms of SETX, which are well documented for this large nuclear helicase involved in R-loop control.