Anti p53 do 1 and anti mdm2 smp14

Cell lysates were electrophoresed through 4–12% SDS-PAGE then transferred to PVDF membranes. The primary antibodies indicated in the figures were incubated with the transferred PVDF membranes in blocking buffer at 4 °C overnight, then washed with PBS, followed with anti-mouse-HRP or anti-rabbit-HRP incubation for 1 h at room temperature. Antibody binding was detected with ECL on PVDF using the syngene chemiluminescent western blot imaging system. The following antibodies were used for immunoblotting: anti-p53 (DO-1) and anti-MDM2 (SMP14) from Santa Cruz; anti-PD-L1(E1L3N), anti-PARP and anti-DR5 from Cell Signaling; anti-IL-6 from Abcam; anti-P21 (Ab-1) from EMD Millipore; and anti-Ran from BD bioscience.

Cells were lysed in RIPA buffer (sigma) containing cocktail protease inhibitors (Roche). Equal amounts of cell lysates were electrophoresed through 4–12% SDS-PAGE then transferred to PVDF membranes. The transferred PVDF membranes were blocked with 5% skim milk for 1 hour at room temperature, then incubated with primary antibodies in the blocking buffer at 4°C overnight. Antibody binding was detected on PVDF with appropriate HRP-conjugated secondary antibodies by the syngene imaging system. The following antibodies were used for immunoblotting: anti-p53 (DO-1) and anti-MDM2 (SMP14) from Santa Cruz; anti-PD-L1 (E1L3N), Anti-MDMX/MDM4 (ab16058) from Abcam; anti-PD1 (D4W2J) from Cell Signaling; anti-P21 (Ab-1) from EMD Millipore and anti-Ran from BD bioscience.