CST, 5737S, Tri-Methyl-Histone H4 (Lys20) (D84D2) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying H4 (Lys20) trimethylate. Validated for WB,ChIP,ChIP,C&R,C&T. Available in 2 sizes. Highly specific and rigorously validated in-house, Tri-Methyl-Histone H4 (Lys20) (D84D2) Rabbit Monoclonal Antibody (CST #5737) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq 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. The CUT&RUN dilution was determined using CUT&RUN Assay Kit # 86652 . The CUT&Tag dilution was determined using CUT&Tag Assay Kit # 77552 . Western Blotting: 1:1000 Chromatin IP: 1:50 Chromatin IP-seq: 1:50 CUT&RUN: 1:50 CUT&Tag: 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: Western Blotting, Chromatin IP, Chromatin IP-seq, CUT&RUN, CUT&Tag Specificity / Sensitivity Tri-Methyl-Histone H4 (Lys20) (D84D2) Rabbit Monoclonal Antibody recognizes endogenous levels of histone H4 protein only when tri-methylated at Lys20. This antibody does not cross-react with non-methylated, mono-methylated or di-methylated histone H4 Lys20. This antibody detects a 95 kDa non-specific protein of unkown origin. 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 H4 in which Lys20 is tri-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 H4; H4 clustered histone 1; H4 histone family, member A; H4-16; H4/A; H4/B; H4/C; H4/D; H4/E; H4/G; H4/H; H4/I; H4/J; H4/K; H4/M; H4/N; H4/O; H4C1; H4C11; H4C12; H4C13; H4C14; H4C15; H4C2; H4C3; H4C4; H4C5; H4C6; H4C8; H4C9; H4F2; H4FA; H4FB; H4FC; H4FD; H4FE; H4FG; H4FH; H4FI; H4FJ; H4FK; H4FM; H4FN; H4FO; HIST1H4A; HIST1H4B; HIST1H4C; HIST1H4D; HIST1H4E; HIST1H4F; HIST1H4H; HIST1H4I; HIST1H4J; HIST1H4K; HIST1H4L; HIST2H4; HIST2H4A; HIST2H4B; HIST4H4; histone 1, H4a; histone cluster 1 H4 family member a; histone cluster 1, H4a; Histone H4 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 11 Source/Isotype: Rabbit IgG