Histone H2a [ac Lys5] Antibody

Western Blot: Histone H2a [ac Lys5] Antibody [NBP2-59261]

Western Blot: Histone H2a [ac Lys5] Antibody [NBP2-59261] - 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 H2AK5ac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (kDa) is shown on the left

Immunocytochemistry/ Immunofluorescence: Histone H2a [ac Lys5] Antibody [NBP2-59261]

Immunocytochemistry/Immunofluorescence: Histone H2a [ac Lys5] Antibody [NBP2-59261] - HeLa cells were stained with the antibody against H2AK5ac 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 H2AK5ac 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 H2a [ac Lys5] Antibody [NBP2-59261] - ChIP assays were performed using human HeLa cells, the antibody against H2AK5ac 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).