Saso2 cells were incubated at 37°C for 24 hours with vehicle control (0 μM) and different concentrations of Amifostine (ab141060). Decreased expression of CDK1 (phospho Y15) (ab133463) in Saso2 cells correlates with an increase in Amifostine concentration, as described in literature.Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab133463 at 1 μg/ml and ab32384 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.