Histone H3 [Trimethyl Lys36] Antibody - BSA Free

Western Blot: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - Histone 3 [Trimethyl Lys36] Antibody [NBP2-54611] - 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 H3K36me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight ~16 kDa.

Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - Histone 3 [Trimethyl Lys36] Antibody [NBP2-54611] - HeLa cells were stained with the antibody against H3K36me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10 min. and blocked with PBS/Triton X-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K36me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Chromatin Immunoprecipitation Sequencing: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - ChIP assays were performed using human K562 cells, the antibody against H3K36me3 and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 100,000 cells. A titration consisting of 0.2, 0.5, 1 and 2 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 for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the coding region of the inactive MB gene and the Sat satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).