Anti p21waf1 cip1 antibody

Total proteins were extracted from cultured cells using RIPA buffer containing protease inhibitor and phosphatase inhibitor (Thermo Fisher Scientific, Waltham, MA, USA). Proteins were subjected to 12% SDS‐PAGE (Bio‐Rad, Hercules, CA, USA). The separated proteins were transferred to Immun‐Blot PVDF membranes (Bio‐Rad) and incubated with anti‐ACTB antibody (A2066; Sigma‐Aldrich; 1:1000 dilution), anti‐cleaved PARP antibody (#5625; Cell Signaling Technology, Beverly, MA, USA; 1:1000 dilution), anti‐p21Waf1/Cip1 antibody (#2947; Cell Signaling Technology; 1:1000 dilution), anti‐Bax antibody (#5023; Cell Signaling Technology, Beverly, MA, USA; 1:1000 dilution), anti‐p53 antibody (Clone DO‐7; DAKO Cytomation, Carpinteria, CA, USA; 1:1000 dilution) and anti‐Noxa antibody (OP180; Calbiochem, Darmstadt, Germany; 1:1000 dilution) at 4°C overnight. Next, the membranes were incubated with HRP‐linked anti‐rabbit IgG (GE Healthcare Biosciences, Piscataway, NJ, USA) and HRP‐linked anti‐mouse IgG (GE Healthcare Biosciences) at a dilution of 1:100 000 for 1 hour at room temperature. The antigen–antibody complex was detected using an ECL Prime Western Blotting Detection Kit (GE Healthcare Biosciences).