Western blot analysis of PLK1 on different lysates with Rabbit anti-PLK1 antibody (HA500281) at 1/1,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 100nM Nocodazole for 24 hours whole cell lysate Lane 3: HeLa treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours whole cell lysate Lane 4: HT-29 whole cell lysate Lane 5: HT-29 treated with 100nM Nocodazole for 24 hours whole cell lysate Lane 6: HT-29 treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours whole cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 68 kDa Observed band size: 68 kDa Exposure time: 2 minutes; 4-20% 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 (HA500281) 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 1:100,000 dilution was used for 1 hour at room temperature.