Western Blot: Histone H4 [Dimethyl Lys20] Antibody [NB21-2089] - Analysis of Histone H4K20me2 in C. elegans embryo lysate.
Immunocytochemistry/ Immunofluorescence: Histone H4 [Dimethyl Lys20] Antibody [NB21-2089] - Histone H4 K20me2 antibody was tested at 1:50 in HeLa cells with FITC (green). Nuclei (Blue) were counterstained with DAPI ...read more
Chromatin Immunoprecipitation: Histone H4 [Dimethyl Lys20] Antibody [NB21-2089] - Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of NB21-2089 and 20ul of magnetic IgG beads per ...read more
Western Blot: Histone H4 [Dimethyl Lys20] Antibody [NB21-2089] - Detection of H4K20me2 in HeLa histone preps.
Dot Blot: Histone H4 [Dimethyl Lys20] Antibody [NB21-2089] - Analysis of H4K20me2 using peptides stated above in 4, 3, 2 and 1 picomoles of peptide.
Chromatin Immunoprecipitation 2-5 ug per million cells
Dot Blot 1:1000
Western Blot 1:2000
In Western blot, a band is seen at ~13kDa. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
When K20 is di-methylated on H4, the downstream effect is silencing of genes. This modification is also necessary and sufficient for 53BP1 binding, which is a prerequisite for DNA repair, a highly conserved mechanism. H4K20me2 is associated with hypoacetylation, and inactivation of certain genes. NSD1 and Suv420h1/2 trigger the dimethylation of K20, but only after SET8 catalyzes the mono-methylation of the same lysine, indicating a complex regulation of this modification that can so seriously affect replication and stability of genomic information.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
FAQs for Histone H4 Antibody (NB21-2089). (Showing 1 - 2 of 2 FAQs).
We have a question about the ChIP result graphs obtained by Epi-Plus antibodies. I am sorry that I could not interpret the results. Could you tell us the interpretation of the results of NB21-2088 and NB21-2089 for example?
The graphs for NB21-2088 and NB21-2089 show the relative amount of DNA for each listed target that was co-precipitated with chromatin. Briefly, cell lysates were incubated with magnetic beads and antibodies for specific histone modifications. The beads and antibody will isolate only those histone complexes that have the specific modification (for example, Dimethyl Lys20 for NB21-2089). These histone complexes are also bound to DNA, therefore any DNA sequences that are associated with these complexes will also be isolated. Once the histones/DNA are immuoprecipitated, the DNA is extracted and qPCR is performed to measure the relative abundance of certain genes. For NB21-2089, the graph shows that genes whose protein is highly expressed in HeLa cells are co-precipitated with NB21-2089 much LESS than genes that are silent in these cell types. This is expected because the modification that NB21-2089 targets is associated with gene silencing, therefore you would expect less GAPDH signal, and more MYOD1 signal (which is not expressed in HeLa cells).
I am wondering what is the immunogen peptide sequence of H4K20me2 Antibody NB21-2089
The immunogen is Synthetic dimethylated peptide surrounding Lysine 20 of human Histone H4 [Swiss Prot P62805]. The exact sequence is considered proprietary.
Histone H4: Implications in Liver Cancer Histones are highly conserved proteins that function in the organization of nuclear DNA to create chromatin in eukaryotic cells. Post-translational alterations of histones are critical to monitoring and regulating DNA structure, expression, and gene t... Read full blog post.
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