Histone extracts (15 μg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) 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.


A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) with peptides containing other modifications of histone H3 and the unmodified H3K9 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.


Mouse NIH3T3 cells were stained with the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) 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 H3K9me2 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit 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.


To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody against H3K9me2 (Cat. No. bs-53118R), crude serum and Flow through. 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 1:103,000.