Trichostain A treated and untreated HeLa cell lysates were subjected to SDS PAGE followed by western blot with 82823-2-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with Alpha Tubulin Monoclonal antibody (66031-1-Ig) as loading control.

Dot blot analysis was used to confirm the specificity of Acetyl-Histone H2A (Lys9) antibody. Acetylated peptides were spotted onto NC and probed with antibody at 1 µg/ml.The amount of peptide (μg/mL) spotted is indicated next to each row. Column 1: H2AK9ac. Column 2: Unmodified H2AK9. Column 3: H2AK5ac. Column 4: Unmodified H2AK5. Column 5: H2AK13Ac. Column 6: Unmodified H2AK13. Column 7: H2AK15ac. Column 8:Unmodified H2AK15. Column 9: Blank(PBS).

Chromatin was prepared from HeLa cells. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 15 µg of cross-linked chromatin, 5 µg of Acetyl-Histone H2A (Lys9) (82823-2-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.