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Specifications

Clone Host Grade Applications Monoclonal Rabbit Affinity Purified B IP IH IC IF

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).
Catalog #	H5110-14J3
Applications	Suitable for use in Immunofluorescence, Western Blot, Immunoprecipitation, ChIP, Immunohistochemistry and Immunocytochemistry. Other applications not tested.
Recommended Dilution	Western Blot: 1:1000
	Immunohistochemistry (paraffin): 1:1500
	Immunofluorescence (IF-IC): 1:800
	Immunoprecipitation: 1:25
	Chromatin IP: 1:50
	Optimal dilutions to be determined by the researcher.
Storage and Stability	May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. 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.
CAS Number	n/a
Clone Type	Monoclonal
Isotype	IgG
Clone No	7H27
Host	Rabbit
Source	Human
Concentration	As reported
Form	Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol and less than 0.02% sodium azide.
Purity	Purified by Protein A and immunoaffinity chromatography.
Immunogen	Synthetic peptide (KLH-coupled) corresponding to the amino terminus of histone H3 in which Lys4 is di-methylated.
Specificity	Detects endogenous levels of 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: Human, monkey, mouse and rat.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.