CST, 33587S, Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit Monoclonal Antibody (ChIP Formulated)

Monoclonal Antibody for studying H3 (Lys36) dimethylate. Validated for Chromatin IP,CUT&RUN. Available in 2 sizes. Highly specific and rigorously validated in-house, Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit Monoclonal Antibody (ChIP Formulated) (CST #33587) is ready to ship. Product Usage Information For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. The CUT&RUN dilution was determined using CUT&RUN Assay Kit # 86652 . Chromatin IP: 1:50 CUT&RUN: 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. Protocol Available protocols: Chromatin IP, CUT&RUN Specificity / Sensitivity Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit Monoclonal Antibody (ChIP Formulated) detects endogenous levels of histone H3.1, histone H3.2, and histone H3.3, only when di-methylated on Lys36. This antibody does not cross-react with non-methylated, mono-methylated, or tri-methylated Lys36. In addition, this antibody does not cross-react with di-methylated histone H3 Lys4, Lys9, Lys27, Lys79 or di-methylated histone H4 Lys20. Species Reactivity: Human Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys36 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 SENSITIVITY: Endogenous MW (kDa): 17 Source/Isotype: Rabbit IgG