Technical Data
H5110-14Z1
Histone H3, dimethyl (Lys4)
Description:
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 acetyla- tion, 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 develop- ment (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 methyltrans- ferases (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 and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000
Immunofluorescence (IF-IC): 1:400
Immunohistochemistry (Paraffin): 1:300. Antigen retrieval: heating in citrate buffer.
Immunoprecipitation: 1:25
Chromatin IP: 1:25
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul-20CBlue IceRabbit
Concentration:
Not Determined
Immunogen:
Synthetic peptide corresponding to the amino terminus of histone H3 in which Lys4 is di-methylated (KLH).
Purity:
Purified by immunoaffinity chromatography.
Form
Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.
Specificity:
Recognizes endogenous levels of histone H3 when di-methylated on Lys4 at ~17kD. Species crossreactivity: human, mouse, monkey and rat. Slight cross-reactivity with mono-methylated Lys4, but does not cross-react with non-methylated or tri-methylated Lys4. Does not cross-react with mono-methylated, di-methylated or tri-methylated histone H3 at Lys9, Lys27, Lys36 or histone H4 at Lys20.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Peterson, C.L. and Laniel, M.A. (2004) Curr. Biol. 14, R546R551. 2. Kubicek, S. et al. (2006) Ernst Schering Res. Found Workshop , 127. 3. Lin, W. and Dent, S.Y. (2006) Curr. Opin. Genet. Dev. 16, 137142. 4. Lee, D.Y. et al. (2005) Endocr. Rev. 26, 147170. 5. Daniel, J.A. et al. (2005) Cell Cycle 4, 919926. 6. Shi, X. et al. (2006) Nature 442, 9699. 7. Wysocka, J. et al. (2006) Nature 442, 8690. 8. Wysocka, J. et al. (2005) Cell 121, 859872. 9. Trojer, P. and Reinberg, D. (2006) Cell 125, 213217.