Histone H3 (K4me3) polyclonal antibody (Cat # PAB15454) was used in ChIP with wild type yeast chromatin (Lane 1) ora strain with a point mutation in the code for Lysine 4 (Lane 2).
Western Blot (Cell lysate)
Acid treated HeLa (10 ug) extract was treated with a 1 : 100 dilution of Histone H3 (K4me3) polyclonal antibody (Cat # PAB15454) prior to visualization.
Dot Blot (Peptide)
To confrm the specifcity of the antisera a dot blot system was used with amounts of peptides from 10-250 pmoles. Unmodifed, monomethylated, dimethylated and trimethylated peptides surrounding the lysine 4 site on Histone H3 were spotted in lanes 1-4 respectively. Lanes 5-8 contained peptides starting at residue 6, with the sequence, TARKSTGGKAPRKQLAT, encompassing the residue at lysine 9 on Histone H3 that is unmodifed, monomethylated, dimethylated and trimethylated respectively. Lanes 9-12 contained peptides encompassing the residue at lysine 27 on Histone H3 that is unmodi-fed, monomethylated, dimethylated and trimethylated respectively.