Histone H3 Antibody (1B1B2)

Western Blot: Histone H3 Antibody (1B1B2) [NBP2-59277]

Western Blot: Histone H3 Antibody (1B1B2) [NBP2-59277] - Whole cell extracts from HeLa cells (40 ug, lane 1) and 1 ug of recombinant histone H3 and H4 (lane 2 and 3). The antibody was diluted 1:5000 in TBS-Tween containing 5% skim milk. Observed molecular weight is ~17 kDa.

Immunocytochemistry/ Immunofluorescence: Histone H3 Antibody (1B1B2) [NBP2-59277]

Immunocytochemistry/Immunofluorescence: Histone H3 Antibody (1B1B2) [NBP2-59277] - HeLa cells were stained with the monoclonal antibody against H3 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa Fluor 594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Chromatin Immunoprecipitation: Histone H3 Antibody (1B1B2) [NBP2-59277] - ChIP assays were performed using human HeLa cells, the monoclonal antibody against H3 and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit for Histones, 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 promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure shows the recovery, expressed as a %input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).