CST, 9728S, Di-Methyl-Histone H3 (Lys27) (D18C8) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying H3 (Lys27) dimethylate. Validated for WB,IP,IF,F,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, Di-Methyl-Histone H3 (Lys27) (D18C8) Rabbit Monoclonal Antibody (CST #9728) is ready to ship. Product Usage Information For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. Western Blotting: 1:1000 Immunoprecipitation: 1:50 Immunofluorescence (Immunocytochemistry): 1:800 - 1:3200 Flow Cytometry (Fixed/Permeabilized): 1:400 - 1:1600 Chromatin IP: 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 27471 . Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP Specificity / Sensitivity Di-Methyl-Histone H3 (Lys27) (D18C8) Rabbit Monoclonal Antibody detects endogenous levels of histone H3 when di-methylated on Lys27. This antibody does show some cross-reactivity with mono-methylated Lys27, but does not cross-react with non-methylated or tri-methylated Lys27. Also, this antibody shows some cross-reactivity with histone H2B when di-methylated on Lys5. In addition, this antibody does not cross-react with mono-methylated, di-methylated or tri-methylated histone H3 Lys4, Lys9, Lys36 or histone H4 Lys20. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys27 is di-methylated. Background 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 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). Alternate Names H3; H3 clustered histone 1; H3 histone family, member A; H3/A; H31; H3C1; H3C10; H3C11; H3C12; H3C2; H3C3; H3C4; H3C6; H3C7; H3C8; H3FA; H3FB; H3FC; H3FC HIST1H3C; H3FD; H3FF; H3FH; H3FI; H3FJ; H3FK; H3FL; HIST1H3A; HIST1H3B; HIST1H3C; HIST1H3D; HIST1H3E; HIST1H3F; HIST1H3G; HIST1H3H; HIST1H3I; HIST1H3J; histone 1, H3a; histone cluster 1 H3 family member a; histone cluster 1, H3a; Histone H3; Histone H3.1; Histone H3/a; Histone H3/b; Histone H3/c; Histone H3/d; Histone H3/f; Histone H3/h; Histone H3/i; Histone H3/j; Histone H3/k; Histone H3/l Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 17 Source/Isotype: Rabbit IgG