Western blot analysis of Cip4 on different lysates with Rabbit anti-Cip4 antibody (HA721029) at 1/1,000 dilution. Lane 1: JAR cell lysate, 10 µg/Lane Lane 2: HepG2 cell lysate, 10 µg/Lane Lane 3: 293T cell lysate, 10 µg/Lane Lane 4: NIH/3T3 cell lysate, 10 µg/Lane Lane 5: Mouse colon tissue lysate, 20 µg/Lane Predicted band size: 68 kDa Observed band size: 80 kDa Exposure time: 2 minutes; 8% 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 (HA721029) 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 1/50,000 dilution was used for 1 hour at room temperature.