Application
Dot blot analysis of peptides containing histone modifications and the unmodified H4K20 using H4K20me3 antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane for analysis.Application
ChIP was performed with K562 extracts and either control IgG (1 μg) or Histone H4K20me3 (tri-Methyl Lys20) antibody (0.2, 0.5, 1 and 2 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls ZNF510 and the Sat2 satellite repeat region, and to negative controls the promoters of the active genes EIF4A2 and GAPDH.Application
ELISA was performed using a serial dilution of Histone H4K20me3 (tri-Methyl Lys20) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:21,700.Application
WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H4K20me3 antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.Application
ICC/IF analysis of HeLa cells using H4K20me3 antibody (green) and DAPI (blue). Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with H4K20me3 antibody at a dilution of 1:500.| Product Name | Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to Histone H4K20me3 (trimethyl Lys20)Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene is intronless and encodes a member of the histone H4 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the histone microcluster on chromosome 6p21.33. [provided by RefSeq, Jul 2008] |
| Immunogen | Polyclonal antibody raised in rabbit against the region of histone H4 containing the trimethylated lysine 20 (H4K20me3), using a KLH-conjugated synthetic peptide. |
| Modification | trimethyl Lys20 |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | DotELISAICC/IFWBChIP |
| ChIP/ChIP-seq (0.5-1 µg/IP), ELISA (1:500), Dot blotting/Peptide array (1:20,000/1:10,000), Western blotting (1:1,00), Immunofluorescence (1:500): |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | H4FM H4M histone cluster 1 H4i HIST1H4I H4/m |
|---|---|
| Target | H4 (tri-Methyl Lys20) |
Application
Dot blot analysis of peptides containing histone modifications and the unmodified H4K20 using H4K20me3 antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane for analysis.
Application
ChIP was performed with K562 extracts and either control IgG (1 μg) or Histone H4K20me3 (tri-Methyl Lys20) antibody (0.2, 0.5, 1 and 2 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls ZNF510 and the Sat2 satellite repeat region, and to negative controls the promoters of the active genes EIF4A2 and GAPDH.
Application
ELISA was performed using a serial dilution of Histone H4K20me3 (tri-Methyl Lys20) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:21,700.
Application
WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H4K20me3 antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.
Application
ICC/IF analysis of HeLa cells using H4K20me3 antibody (green) and DAPI (blue). Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with H4K20me3 antibody at a dilution of 1:500.| Application Notes | ChIP/ChIP-seq (0.5-1 µg/IP), ELISA (1:500), Dot blotting/Peptide array (1:20,000/1:10,000), Western blotting (1:1,00), Immunofluorescence (1:500): |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide and 0.05% ProClin 300. |
