Western blot analysis of alpha 1 Spectrin on different lysates with Rabbit anti-alpha 1 Spectrin antibody (HA500357) at 1/1,000 dilution. Lane 1: K562 cell lysate Lane 2: NIH/3T3 cell lysate Lane 3: A549 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 280 kDa Observed band size: 280 kDa Exposure time: 2 minutes; 6% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500357) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.