Histone H3 [Dimethyl Lys79] Antibody

Chromatin Immunoprecipitation: Histone H3 [Dimethyl Lys79] Antibody [NBP2-59194] - ChIP assays were performed using human HeLa cells, the antibody against H3K79me2 and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1) gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Western Blot: Histone H3 [Dimethyl Lys79] Antibody [NBP2-59194]

Western Blot: Histone H3 [Dimethyl Lys79] Antibody [NBP2-59194] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K79me2 diluted 1:200 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.

Dot Blot: Histone H3 [Dimethyl Lys79] Antibody [NBP2-59194] - Analysis was performed to test the cross reactivity of the Diagenode antibody against H3K79me2 with peptides containing other modifications and unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.