GRP78/BIP/HSPA5 Antibody

Western blot analysis of GRP78/BIP/HSPA5 using anti-GRP78/BIP/HSPA5 antibody. 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 ug of sample under reducing conditions. Lane 1: human placenta tissue lysates, Lane 2: human U-87MG whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human T-47D whole cell lysates, Lane 5: rat liver tissue lysates, Lane 6: rat brain tissue lysates, Lane 7: mouse liver tissue lysates, Lane 8: 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-GRP78/BIP/HSPA5 antigen affinity purified polyclonal antibody 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 1:5000 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 GRP78/BIP/HSPA5 at approximately 78 kDa. The expected band size for GRP78/BIP/HSPA5 is at 78 kDa.