Anti acetyl p53 k305 antibody

ESCs or EBs were lysed with RIPA buffer (Pierce) in the presence of EDTA free protease inhibitor cocktail (Sigma). Histone extraction on EBs was performed using a histone extraction kit (Abcam) as described previously (Khan et al., 2015 (link)). Protein concentration was determined using Pierce BCA protein assay kit (ThermoFisher Scientific). 1.0 μg of histone extracts, or 10 μg of whole cell lysate was resolved on 4%–20% precast gel (Bio-Rad) was transferred to 0.45 μm PVDF membrane (Millipore). For immunoprecipitation, day 2 WT and Wdr5^KO EBs were harvested and lysed in RIPA buffer with EDTA free protease inhibitor cocktail (Sigma). 1 mg sonicated cell lysates were subjected to immunoprecipitation using WDR5 antibody (Bethyl). The following primary antibodies were used for probing: anti-HA (1:10,000, Abcam, ab9110), anti-WDR5 (1:5000, R&D), anti-WDR5 (1:5,000, Bethyl), anti-Flag (1:2,000, Sigma), anti-H3 (1:10,000, Abcam), anti-H3K4Me1 (1:5,000, Millipore), anti-H3K4Me2 (1:10,000, Millipore), anti-H3K4Me3 (1:10,000, Abcam), anti-P53 (1:5,000, Cell Signaling), anti-Tubulin (1:10,000, Cell signaling), anti-β-Actin (1:10,000, Cell signaling), anti-Phospho-p53 (Ser15) Antibody (1:5,000, Cell Signaling), anti-Phospho-p53 (Ser392) Antibody (1:5,000, Abcam), anti-Acetyl-p53 (K305) Antibody (1:5,000, Abcam), anti-Actyl-p53 (K379) Antibody (1:5,000, Cell Signaling).