Figure 1. Western blot analysis of HSPA4 using anti-HSPA4 antibody (A03618-2). 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 293T whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: rat brain tissue lysates, Lane 4: rat C6 whole cell lysates, Lane 6: mouse brain tissue lysates, Lane 7: mouse NIH/3T3 whole cell lysates, Lane 8: mouse Raw264.7 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-HSPA4 antigen affinity purified polyclonal antibody (Catalog # A03618-2) at 0.25 microg/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-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for HSPA4 at approximately 110 kDa. The expected band size for HSPA4 is at 96,14 kDa.