Western blot analysis of Doppel in Lane 1: human SHG-44 whole cell lysate, Lane 2: rat testis tissue lysates, Lane 3: rat kidney tissue lysates, Lane 4: mouse testis tissue lysates, Lane 5: mouse kidney tissue lysates, Lane 6: mouse brain tissue lysates. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with Doppel polyclonal antibody (Product # PA5-95577) using a 0.5 µg/mL dilution, followed by a goat anti-rabbit IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).