Technical Data

H5110-14J3
Clone Type
Monoclonal
Host
Rabbit
Source
Human
Isotype
IgG
Clone Number
7H27(C64G9)
Grade
Affinity Purified
Applications
ChIP IF IHC IP WB
Crossreactivity
Hu Mk Mo Rt
Gene ID
8350
Shipping Temp
Blue Ice
Storage Temp
-20°C
Rabbit Anti-Histone H3, Dimethyl (Lys4)

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 have 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 recent discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2 and JHDM1 has shown that methylation is a reversible epigenetic mark (9).

Applications
Suitable for use in Immunofluorescence, Western Blot, Immunoprecipitation, ChIP, Immunohistochemistry. Other applications not tested.
Recommended Dilution
Western Blot: 1:1000, incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight. Immunohistochemistry (paraffin): 1:1500 Immunofluorescence (IF-IC): 1:800 Immunoprecipitation: 1:25 ChIP: 1:50 Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic peptide corresponding to the N-terminus of human histone H3 in which Lys4 is di-methylated.
Form
Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 0.02% sodium azide, 50% glycerol.
Purity
Purified by Protein A and peptide affinity chromatography.
Specificity
Detects endogenous levels of human histone H3 only when di-methylated on Lys4. Does not crossreact with non-methylated, mono-methylated, or tri-methylated Lys4. In addition, does not crossreact with di-methylated or tri-methylated histone H3 Lys9, Lys27, Lys36 or histone H4 Lys20. Species Crossreactivity: monkey, mouse and rat.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1 Peterson, C.L. and Laniel, M.A. (2004) Curr. Biol. 14, R546-R551. 2 Kubicek, S. et al. (2006) Ernst Schering Res. Found Workshop , 1-27. 3 Lin, W. and Dent, S.Y. (2006) Curr. Opin. Genet. Dev. 16, 137-142. 4 Lee, D.Y. et al. (2005) Endocr. Rev. 26, 147-170. 5 Daniel, J.A. et al. (2005) Cell Cycle 4, 919-926. 6 Shi, X. et al. (2006) Nature 442, 96-99. 7 Wysocka, J. et al. (2006) Nature 442, 86-90. 8 Wysocka, J. et al. (2005) Cell 121, 859-872. 9 Trojer, P. and Reinberg, D. (2006) Cell 125, 213-217.
USBio References
No references available
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