Non-treated PC-3 and MMS treated PC-3 cells were subjected to SDS PAGE followed by western blot with 29012-1-AP (Phospho-CHEK2 (Thr68) antibody) at dilution of 1:1000 incubated at room temperature for 4℃ overnight. The membrane was stripped and re-blotted with GAPDH antibody as loading control.