Histone H4 [Trimethyl Lys20] Antibody

Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using Histone H4 [Trimethyl Lys20] Antibody 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 (in kDa) is shown on the left.

Immunocytochemistry/ Immunofluorescence: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259]

Immunocytochemistry/Immunofluorescence: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - (Top): Human osteosarcoma (U2OS) cells were stained with and DAPI. Cells were fixed with ice cold methanol for 10' and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 6A: cells were immunofluorescently labeled with (left) diluted 1:300 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. (Bottom): Staining of the cells with the after incubation of the antibody with blocking peptide, concentration: 5 ng/ul.

Chromatin Immunoprecipitation: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - ChIP assays were performed using undifferentiated human teratocarcinoma cells (NCCIT), the antibody against H4K20me3 and optimized PCR primer sets for qPCR. Sheared chromatin from 10,000 cells was used per ChIP experiment. The antibody was tested at two different dilutions of 1:500 and 1:100. The pre-immune serum (PI) was used as a negative control. Quantitative PCR was performed using primer sets for the satellite repeat Sat2 as a positive control and for the promoter of the house keeping gene c-fos, as a negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H4K20me3 is preferably present at heterochromatin.