Histone H3 [Monomethyl Lys4] Antibody - BSA Free

Western Blot: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - 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 H3K4me1. The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.

Immunocytochemistry/Immunofluorescence: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - HeLa cells were stained with the antibody against H3K4me1 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 H3K4me1 antibody (left) diluted 1:200 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 H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - ChIP assays were performed using human K562 cells, the antibody against H3K4me1 and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration consisting of 1, 2, 5 and 10 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 a region surrounding the ACTB and GAS2L1 gene, respectively, used as positive controls, and for the promoters of the GAPDH and EIF4A2 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).