Western blot analysis of RNF146 in different various lysates. Lane 1: SH-SY5Y cell lysate; Lane 2: NIH/3T3 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 39 kDa. Observed band size: 39 kDa. Exposure time: 2 minutes; 10% SDS-PAGE gel. Primary antibody RNF146 recombinant monoclonal antibody (Product # MA5-50800) with a dilution of 1:1,000 was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody with a dilution of 1:300,000 was used for 1 hour at room temperature. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature.