ChIP was performed with 1 μg of the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) as described above. The IP’d DNA was subsequently analyzed on an Illumina HiSeq 2000. Library preparation, cluster generation, and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the peak distribution along the complete sequence and a 5 Mb region of chromosome 1 (figure A and B) and in two 300 kb regions surrounding the EIF2S3 and CCT5 positive control genes (figure C and D).


To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K79me2 (Cat. No. bs-53111R). 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 1:6,600.


A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) with peptides containing other modifications and unmodified sequences of histone H3. 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 1:5,000. The figure shows a high specificity of the antibody for the modification of interest.


Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) diluted 1:200 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.