Anti p p38 sc 7973

The cells were treated as above, harvested, washed with ice-cold PBS, and lysed with ice-cold RIPA lysis buffer. Protein concentrations were measured using the BCA Protein Assay Reagent (Pierce, Rockford, IL, USA). Equivalent amounts of protein were separated by sodium dodecyl sulfate-polyacrylamide electrophoresis in 8%–12% gels and transferred to polyvinylidene difluoride membranes (Millipore Corp., Billerica, MA, USA). The membranes were blocked with 6% nonfat dry milk at room temperature for 1 hour, and incubated with the primary antibodies overnight at 4°C. The following primary antibodies were used: anti-Bax (ab10813; Abcam), anti-Bcl-2 (ab7973; Abcam), anti-p-Erk1/2 (Thr202/Tyr204) (sc-16982; Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-p38 (sc-7973; Santa Cruz Biotechnology), anti-P-p53 (S15) (12571S; Cell Signaling, Danvers, MA, USA), anti-RAD51 (ab88572; Abcam), and anti-β-actin (ab8227; Abcam). The membranes were washed and incubated with horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 hour. Blots were developed using enhanced chemiluminescence.