Figure 1. Western blot analysis of SHC using anti-SHC antibody (PB9391). 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 50ug of sample under reducing conditions. Lane 1: human A431 whole cell lysate, Lane 2: human Hela whole cell lysate, Lane 3: human HepG2 whole cell lysate, Lane 4: human Jurkat whole cell lysate, Lane 5: human K562 whole cell lysate, Lane 6: human THP-1 whole cell lysate, Lane 7: rat C6 whole cell lysate, Lane 8: mouse thymus tissue lysate, Lane 9: mouse RAW246.7 whole cell lysate, Lane 10: mouse NIH3T3 whole cell lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-SHC antigen affinity purified polyclonal antibody (Catalog # PB9391) at 0.5 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-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. Specific bands were detected for SHC at approximately 46, 52, 66KD. The expected band size for SHC are at 46, 52, 66KD.