Western blot analysis of CHRNA3 in Lane 1: human HeLa whole cell lysate, Lane 2: human MDA-MB-453 whole cell lysate, Lane 3: human Jurkat whole cell lysate, Lane 4: human HepG2 whole cell lysate, Lane 5: human SK-OV-3 whole cell lysate, Lane 6: human PANC-1 whole cell lysate, Lane 7: mouse thymus tissue lysate using 50 µg (reducing conditions) per well. Electrophoresis was performed on 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours and protein was transferred to a nitrocellulose membrane at 150mA for 50-90 minutes. Sample was blocked with 5% Non-fat Milk/TBS for 1.5 hours at room temperature, incubated with CHRNA3 polyclonal antibody (Product # PA5-79045) at a dilution of 0.5 µg/mL (overnight at 4°C), followed by goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10,000. Signal development was performed using a chemiluminescence (ECL) kit.