Acid treated HeLa extract was treated with a 1 : 500 dilution of antibody Histone H3 (K4me1) polyclonal antibody (Cat # PAB15452) 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. Lane 5 was also spotted with the monomethylated Lys4 peptide. Lanes 6-9 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.