ChIP was performed with 1 μg of the Bioss antibody against H3K9ac (Cat. No. bs-53092R) and the IP’d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer’s 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.


To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K9ac (Cat. No. bs-53092R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:31,700.


To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K9ac (Cat. No. bs-53092R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:31,700.


A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K9ac (Cat. No. bs-53092R) with peptides containing other histone modifications and the unmodified H3K9 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest. Please note that this antibody recognizes the H3K9 acetylation, both in the presence and the absence of the K14 acetylation.


Histone extracts of HeLa cells (15 μg) were analyzed by Western blot using the Bioss antibody against H3K9ac (Cat. No. bs-53092R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.


HeLa cells were stained with the Bioss antibody against H3K9ac (Cat. No. bs-53092R) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K9ac antibody (left) diluted 1:500 in blocking solution followed by secondary antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.