Western blot analysis of TRIP/TRAIP using TRIP/TRAIP antibody (Product # PA5-143982). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. Lane 1: human Daudi whole cell lysates; Lane 2: human U251 whole cell lysates; Lane 3: rat brain tissue lysates; Lane 4: mouse brain 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-TRIP/TRAIP antigen affinity purified polyclonal antibody (Product # PA5-143982) at 0.25 μg/mL overnight at 4°C, 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 1:5,000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for TRIP/TRAIP at approximately 53 kDa. The expected band size for TRIP/TRAIP is at 53 kDa.