Figure 1. Western blot analysis of ATP1A1 using anti-ATP1A1 antibody (PB9482). 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: rat brain tissue lysates, Lane 2: rat brain tissue lysates, Lane 3: mouse brain tissue lysates, Lane 4: mouse brain tissue lysates. 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-ATP1A1 antigen affinity purified polyclonal antibody (Catalog # PB9482) 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-DyLight 647 Conjugated secondary antibody (Left) at a dilution of 1:2000 or a goat anti-rabbit IgG-HRP Conjugated secondary antibody (Right) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ATP1A1 approximately 100 kDa. The expected band size for ATP1A1 is at 74, 110, 113 kDa.