Histone H4 [Dimethyl Lys20] Antibody (C.15200220) - BSA Free

Western Blot: Histone H4 [Dimethyl Lys20] Antibody (C.15200220) [NBP2-59246] - Histone extracts (15 ug) from HeLa cells were analyzed using the antibody against H4K20me2 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (kDa) is shown on the left.

Immunocytochemistry/Immunofluorescence: Histone H4 [Dimethyl Lys20] Antibody (C.15200220) [NBP2-59246] - HeLa cells were stained with the antibody against H4K20me2 and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H4K20me2 antibody (middle) diluted 1:1.000 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 H4 [Dimethyl Lys20] Antibody (C.15200220) [NBP2-59246] - ChIP assays were performed using HeLa cells, the monoclonal antibody against H4K20me2 and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit on the IP-Star automated system, using sheared chromatin from 1 million cells. Respectively 1 and 5 ug of antibody was used per ChIP experiment. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the GAPDH and EIF4A2 genes, used as negative controls, and for the Sat2 and SatA satellite repeats, used as positive controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).