Pan acetylated lysine antibody

Murine TEAD1 residues 211–427, containing the YAP and VGLL4 binding domain, was fused to a polyhistidine tag, expressed in bacteria, and purified on Ni-nitrilotriacetic acid agarose (Qiagen). Peptides VGLL4-TDU-V5 and VGLL4-TDU-KAc-V5 (Figure S3) were chemically synthesized. VGLL4-TEAD1 binding affinity was determined using photonic crystal nanobeam sensors (Yang et al., 2014 ).
Cell or tissue-soluble protein extracts for coIP were immunoprecipitated using protein A or streptavidin Dynabeads (Life Technologies). Acetylation was analyzed using pan-acetylated lysine antibody (Cell Signaling Technology, 9441S) or mass spectrometry (Harvard Medical School Taplin Biological Mass Spectrometry Facility). Murine VGLL4-K216Ac specific antibody was raised in rabbits and isolated by affinity purification.

Lysine acetylation was detected using a pan-acetylated lysine antibody (Cell Signaling #9814). Total Histone H3, as a loading control, was detected using a Histone H3 antibody (Abcam #ab46765). Both antibodies were used at 1:5000 and incubated at 4°C overnight.