Tri-Methyl-Histone H3 (Lys4) AntibodyProduct information
Product Pathways - Chromatin Regulation / Epigenetics
Tri-Methyl-Histone H3 (Lys4) Antibody #9727
|9727L||300 µl (30 western blots)||---||In Stock||---|
|9727S||100 µl (10 western blots)||---||In Stock||---|
|9727||carrier free and custom formulation / quantity||email request|
|W||1:1000||Human, Mouse, Rat, Monkey||17||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Xenopus, Zebrafish.
Directions For Use
For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Specificity / Sensitivity
Tri-Methyl-Histone H3 (Lys4) Antibody detects endogenous levels of histone H3 when tri-methylated on Lys4. This antibody shows some cross-reactivity with histone H3 that is di-methylated on Lys4, but does not cross-react with non-methylated or mono-methylated histone H3 Lys4. In addition, the antibody does not cross-react with methylated histone H3 Lys9, Lys27, Lys36 or methylated histone H4 Lys20.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which lysine 4 is tri-methylated. Antibodies are purified by protein A and peptide affinity chromatography.
Tri-Methyl-Histone H3 (Lys4) Antibody specificity was determined by peptide ELISA. Each graph depicts a titration of this antibody and the corresponding reactivity toward the non-methyl, mono-methyl, di-methyl and tri-methyl states of the indicated histone H3 or H4 lysine residue.
Western blot analysis of lysates from HeLa, NIH/3T3, C6 and COS cells, using Tri-Methyl-Histone H3 (Lys4) antibody.
Immunohistochemical analysis of paraffin-embedded human colon using Tri-Methyl-Histone H3 (K4) Rabbit Antibody in the presence of non-methyl peptide (left) or K4 tri-methyl peptide (right).
Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Tri-Methyl-Histone H3 (Lys4) Antibody (green, left) compared to an isotype control (right). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Tri-Methyl-Histone H3 (Lys4) Antibody or 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 GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
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, R546-51.
- Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop , 1-27.
- Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
- Lee, D.Y. et al. (2005) Endocr Rev 26, 147-70.
- Daniel, J.A. et al. (2005) Cell Cycle 4, 919-26.
- Shi, X. et al. (2006) Nature 442, 96-9.
- Wysocka, J. et al. (2006) Nature 442, 86-90.
- Wysocka, J. et al. (2005) Cell 121, 859-72.
- Trojer, P. and Reinberg, D. (2006) Cell 125, 213-7.
- Song, N. et al. (2011) Acta Histochem Cytochem 44, 183-90. Applications: IHC-P (paraffin).
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