Histone H3 [Trimethyl Lys9] Antibody - BSA Free

Western Blot: Histone H3 [Trimethyl Lys9] Antibody [NBP2-59198]

Western Blot: Histone H3 [Trimethyl Lys9] Antibody [NBP2-59198] - 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 H3K9me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~18 kDa.

Immunocytochemistry/ Immunofluorescence: Histone H3 [Trimethyl Lys9] Antibody [NBP2-59198]

Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys9] Antibody [NBP2-59198] - HeLa cells were stained with the Diagenode antibody against H3K9me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me3 antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.

Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys9] Antibody [NBP2-59198] - ChIP assays were performed using human HeLa cells, the antibody against H3K9me3 and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using the "Auto Histone ChIP-seq" kit on the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. QPCR was performed with primers for the heterochromatin marker Sat2 and for the ZNF510 gene, used as positive controls, and for the promoters of the active c-fos and GAPDH genes, 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).