ChIP assays were performed using HeLa cells, H3K9ac Polyclonal Antibody (Cat. No. bs-53092R) and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that acetylation of K9 at histone H3 is associated with the promoters of active genes.

ChIP was performed with 1 \u03bcg of the Bioss antibody against H3K9ac (Cat. No. bs-53092R) and the IP\u2019d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the peak distribution along the complete sequence and an 800 kb region of the X-chromosome (figure A and B) and in 100 kb regions surrounding the GAPDH and EIF4A2 positive control genes ( figure C and D). These results clearly show an enrichment of H3K9ac at the promoters of active genes.