Product Pathways - Chromatin Regulation / Epigenetics
Pan-Methyl-Histone H3 (Lys9) Antibody #4069
|W IP IF-IC ChIP||H M R Mk Z||Endogenous||17||Rabbit|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey Z=Zebrafish
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Pan-Methyl-Histone H3 (Lys9) Antibody detects endogenous levels of histone H3 only when mono-, di-, or tri-methylated on Lys9. The antibody does not cross-react with histone H3 mono-methylated, di-methylated or tri-methylated on Lys27.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which lysine 9 is di-methylated. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of whole cell lysates from HeLa, NIH/3T3, C6 and COS cells, using Pan-Methyl-Histone H3 (Lys9) Antibody.
DAPI staining (left) and immunofluorescent analysis (right) of NIH/3T3 cells, using Pan-Methyl-Histone H3 (Lys9) Antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 μl of Pan-Methyl-Histone H3 (Lys9) Antibody or 2 μl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human AFM Intron 1 Primers #5098, SimpleChIP® Human α Satellite Repeat Primers #4486, SimpleChIP® Human RPL30 Exon 3 Primers #7014, and SimpleChIP® Human GAPDH Exon 1 Primers #5516. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Pan-Methyl-Histone H3 (Lys9) Antibody specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated di-methyl histone H3 (Lys9) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the mono-, di- and tri-methyl histone H3 (Lys9) peptides competed away binding of the antibody.
The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1 has shown that methylation is a reversible epigenetic marker (9).
- Peterson, C.L. and Laniel, M.A. (2004) Curr. Biol. 14, R546-R551.
- Kubicek, S. et al. (2006) Ernst Schering Res. Found Workshop, 1-27.
- Lin, W. and Dent, S.Y. (2006) Curr. Opin. Genet. Dev. 16, 137-142.
- Lee, D.Y. et al. (2005) Endocr. Rev. 26, 147-170.
- Daniel, J.A. et al. (2005) Cell Cycle 4, 919-926.
- Shi, X. et al. (2006) Nature 442, 96-99.
- Wysocka, J. et al. (2006) Nature 442, 86-90.
- Wysocka, J. et al. (2005) Cell 121, 859-872.
- Trojer, P. and Reinberg, D. (2006) Cell 125, 213-217.
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- 9002 SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
- 9003 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9007 ChIP-Grade Protein G Agarose Beads
- 9006 ChIP-Grade Protein G Magnetic Beads
- 2729 Normal Rabbit IgG
- 9701 Phospho-Histone H3 (Ser10) Antibody
- 9753 Di-Methyl-Histone H3 (Lys9) Antibody
- 9715 Histone H3 Antibody
- 2616 HP1α Antibody
- 2613 HP1β Antibody
- 2619 HP1γ Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.