ChIP was performed with the nuclear extract of WT or H3K4R mutant S. pombe cells and either 0.5 μg of H3K4ac antibody or control rabbit IgG. The precipitated DNA was detected by QPCR with primer set targeting to adh1 or the pericentric repeat regions and for the euchromatic adh1.

Dot blot analysis of peptides containing histone H3 modifications and the unmodified H3K4 sequence using H3K4ac antibody at a dilution of 1:10,000. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane for analysis.

ICC/IF analysis of wild type and H3K4R mutant S. pombe cells using H3K4ac antibody (in red) and by Hoechst staining (in blue, left). The antibody was used at a dilution of 1:300.

WB analysis of histone extracts (15 μg) from HeLa cells using H3K4ac antibody diluted 1:500 in TBS-Tween containing 5% BSA.

ELISA was performed using a serial dilution of Histone H3K4ac (acetyl Lys4) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:27,800.