Rabbit Histone H3, dimethyl Monoclonal Antibody | anti-H3 antibody

Histone H3, dimethyl (Lys9)

Cat. Num. AAA606714

• Reactivity: Human, Monkey, Mouse, Rat
• Applications: Western Blot, Immunoprecipitation, Immunofluorescence
• Synonyms: Histone H3; dimethyl; Monoclonal Antibody; dimethyl (Lys9); Anti -Histone H3; anti-H3 antibody

• Host: Rabbit
• Reactivity: Human, Monkey, Mouse, Rat
• Clonality: Monoclonal
• Isotype: IgG
• Clone Number: 10E30
• Specificity: Recognizes human Di-Methyl-Histone H3 (Lys9) (D85B4). Species crossreactivity
• Form/Format: Supplied in liquid 10mM sodium HEPES pH 7.5, 150mM NaCl, 100ug/ml BSA, 50% glycerol and 0.02% sodium azide.

• Applicable Applications for anti-H3 antibody: Western Blot (WB), Immunoprecipitation (IP), Immunofluorescence (IF)
• Application Notes: Suitable for use in Western Blot, Immunoprecipitation, Immunofluorescence (IF-IC) and Immunoprecipitation (Chromatin).; Dilution: Western Blot: 1:1000. Incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween-20 at 4 degree C with gentle shaking, overnight.; Immunoprecipitation: 1:50; Immunofluorescence (IF-IC) 1:200; Chromatin Immunoprecipitation (CHIP): 1:25
• Immunogen: Synthetic peptide corresponding to the amino terminus of histone H3 in which Lys9 is di-methylated (KLH- coupled).
• Preparation and Storage: May be stored at 4 degree C for short-term only. For long-term storage, aliquot and store at -20 degree C. Aliquots are stable for at least 12 months at -20 degree 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.

Western Blot (WB)

(Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys9) mAb.)

Immunofluorescence (IF)

(Confocal immunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys9) mAb(green). Actin filaments have been labeled with DY-554 phalloidin (red).)

ELISA (EIA)

(Di-Methyl Histone H3 (Lys9) mAb 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 di-methyl histone H3 (Lys9) peptide competed away binding of the antibody.)

Immunoprecipitation (IP)

(Chromatin immunoprecipitations were performed with cross-linked chromatin from 4x10e6 HeLa cells and either 20ul of MBS606714 or 2ul of Normal Rabbit IgG using SimpleChIP Enzymatic Chromatin IP Kit (Magnetic Beads). The enriched DNA was quantified by Real-Time PCR using primers specific for the transcriptionally active GAPDH and RPL30 genes, the inactive AFM gene, and the heterochromatic alpha Satellite repeat element. 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.)

Related Product Information for anti-H3 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 recent discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2 and JHDM1 has shown that methylation is a reversible epigenetic mark (9).

Product Categories/Family for anti-H3 antibody

Antibodies; Abs to Histones

NCBI and Uniprot Product Information

• NCBI GI #: 47551065
• NCBI GeneID: 373335
• NCBI Accession #: NP_999709.1
• NCBI GenBank Nucleotide #: NM_214544.1
• NCBI Official Full Name: histone H3
• NCBI Official Symbol: H3
• NCBI Protein Information: histone H3
• Protein Family: Tyrosine phosphatase