Anti phospho p53 ab

The cells were lysed using RIPA lysis buffer (Nacalai Tesque) added together with a protease and phosphatase inhibitor cocktail (Nacalai Tesque). Equal amounts of proteins (25 µg) were loaded onto the gels (7.5, 10 and 15% gels were used), separated by SDS-PAGE, and then transferred onto Immobilon-P membranes (Millipore Corp.). These membranes were probed with primary antibodies, such as anti-p53 antibody (Ab) (sc-126, 1/1,000), anti-β-actin Ab (sc-47778, 1/1,000), anti-HDAC6 Ab (sc-11420, 1/1,000), anti-acetylated α-tubulin Ab (sc-23950, 1/1,000), and anti-α-tubulin Ab (sc-5286, 1/1,000) were purchased from Santa Cruz Biotechnology, Inc. Anti-PARP Ab (#9542S, 1/1,000), anti-caspase3 Ab (#9665S, 1/1,000), anti-phospho-p53 Ab (#9286, 1/1,000), anti-p21 Ab (#2947S, 1/1,000), anti-phospho-ATR Ab (#9947, 1/1,000), anti-ATR Ab (#2790, 1/1,000), anti-phospho-Chk1 (Ser345) Ab (#2348, 1/1,000), anti-Chk1 Ab (#2360, 1/1,000), anti-phospho-Chk2 (Thr68) Ab (#2197, 1/1,000), anti-Chk2 Ab (#3440, 1/1,000), anti-phospho-Cdc2 (Tyr15) Ab (#4539, 1/1,000), anti-Cdc2 Ab (#9116, 1/1,000), anti-H2A.X Ab (#7631, 1/1,000), and anti-phospho-histone H2A.X (Ser139) Ab (#9718, 1/1,000) were purchased from Cell Signaling Technology, Inc.