Histone H2B [ac Lys15] Antibody

Western Blot: Histone H2B [ac Lys15] Antibody [NBP2-59272]

Western Blot: Histone H2B [ac Lys15] Antibody [NBP2-59272] - 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 H2BK15ac. The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The marker (kDa) is shown on the left.

Immunocytochemistry/ Immunofluorescence: Histone H2B [ac Lys15] Antibody [NBP2-59272]

Immunocytochemistry/Immunofluorescence: Histone H2B [ac Lys15] Antibody [NBP2-59272] - HeLa cells were stained with the antibody against H2BK15ac and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/ TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2BK15ac 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: Histone H2B [ac Lys15] Antibody [NBP2-59272] - ChIP assays were performed using human HeLa cells, the antibody against H2BK15ac and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of 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).