Histone extracts of HeLa cells (15 μg) were analyzed by Western blot using the Bioss antibody against H3K4me2 (Cat. No. bs-53105R) 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 H3K4me2 (Cat. No. bs-53105R) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K4 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. Figure shows a high specificity of the antibody for the modification of interest.


To determine the titer, an ELISA was performed using a serial dilution of H3K4me2 Polyclonal Antibody (bs-53105R), crude serum and flow through 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 1:12,600.


ChIP was performed as described above using 1 μg of the Bioss antibody against H3K4me2 (Cat. No. bs-53105R). The IP’d DNA was 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 X-chromosome (figure A) and in 3 chromosomal regions surrounding the GAPDH, c-fos and ACTB genes (figure B, C, and D respectively).