WB was performed as previously described [50 (link)]. ACTIN was used as a loading control. The following primary antibodies were used: TC2N rabbit polyclonal antibody (1:500; Abcam), p53 mouse monoclonal antibody (1:1000; Santa Cruz Biotechnology), p21 mouse monoclonal antibody (1:1000; Santa Cruz Biotechnology), BAX mouse monoclonal antibody (1:1000; Santa Cruz Biotechnology), Bcl-2 mouse monoclonal antibody (1:1000; Santa Cruz Biotechnology), Cdk5 mouse monoclonal antibody (1:1000; Santa Cruz Biotechnology), phosphor-p53 (Ser-15) rabbit polyclonal antibody (1:500; Bioss), phosphor-p53 (Ser-33) rabbit polyclonal antibody (1:500; Bioss), phosphor-p53 (Ser-46) rabbit polyclonal antibody (1:500; Bioss), ubiquitin rabbit polyclonal antibody (1:500; Abcam) and ACTIN monoclonal antibody (1:2000; Sigma).
For co-immunoprecipitation, cell lysates were prepared and immunoprecipitated with indicated antibody or mouse IgG using the Co-IP Kit (Pierce, Rockford, IL, USA) according to the manufacturer’s instructions. The precipitated protein was assessed by WB using indicated antibody.
For co-immunoprecipitation, cell lysates were prepared and immunoprecipitated with indicated antibody or mouse IgG using the Co-IP Kit (Pierce, Rockford, IL, USA) according to the manufacturer’s instructions. The precipitated protein was assessed by WB using indicated antibody.
Free full text: Click here
