CST, 3306T, G9a/EHMT2 (C6H3) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying G9a. Validated for Western Blotting,Immunofluorescence (Immunocytochemistry),Chromatin IP. Available in 2 sizes. Highly specific and rigorously validated in-house, G9a/EHMT2 (C6H3) Rabbit Monoclonal Antibody (CST #3306) 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 Immunofluorescence (Immunocytochemistry): 1:50 Chromatin IP: 1:100 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, Immunofluorescence (Immunocytochemistry), Chromatin IP Specificity / Sensitivity G9a/EHMT2 (C6H3) Rabbit Monoclonal Antibody detects endogenous levels of total G9a/EHMT2 protein (both the 165 kDa G9a-L and 140 kDa G9a-S isoforms). This antibody does not cross-react with other histone methyltransferases, including GLP. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human G9a/EHMT2 protein. Background G9a, also known as Euchromatic histone-lysine N-methyltransferase 2 (EHMT2), is a member of a family of histone lysine methyltransferases, each of which contains a conserved catalytic SET domain originally identified in Drosophila Su[var]3-9, Enhancer of zeste, and Trithorax proteins (1). Recombinant G9a can mono-, di- and tri-methylate histone H3 on Lys9 and Lys27 (1,2). However, G9a forms a complex with GLP, a G9a-related histone methyltransferase, and together these proteins function as the major euchromatic histone H3 Lys9 mono- and di-methyltransferases, creating transcriptionally repressive marks that facilitate gene silencing (3,4). G9a methylates itself on Lys165, a modification that regulates the association of HP1 repressor proteins with the G9a/GLP complex (5). The G9a/GLP complex also contains Wiz, a zinc finger protein that is required for G9a/GLP hetero-dimerization and complex stability (6). Wiz contains two CtBP co-repressor binding sites, which mediate the association of the G9a/GLP with the CtBP co-repressor complex (6). In addition, G9a and GLP are components of other large transcriptional co-repressor complexes, such as those involving E2F6 and CDP/cut (7-9). G9a interacts with DNMT1, and both proteins are required for methylation of DNA and histone H3 (Lys9) at replication foci, providing a functional link between histone H3 Lys9 and CpG methylation during DNA replication (10). G9a activity is critical for meiotic prophase progression, as mutant mice deficient in germ line G9a show a large loss of mature gametes (11). In addition, G9a facilitates increased global levels of di-methyl histone H3 (Lys9) during hypoxic stress and increased G9a expression is associated with hepatocelluar carcinoma (12,13). Alternate Names ankyrin repeat-containing protein; BAT8; C6orf30; EHMT2; euchromatic histone lysine methyltransferase 2; Euchromatic histone-lysine N-methyltransferase 2; G9A; G9A histone methyltransferase; GAT8; H3-K9-HMTase 3; Histone H3-K9 methyltransferase 3; Histone-lysine N-methyltransferase EHMT2; histone-lysine N-methyltransferase, H3 lysine-9 specific 3; HLA-B associated transcript 8; HLA-B-associated transcript 8; KMT1C; Lysine N-methyltransferase 1C; NG36; Protein G9a Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 160,180 Source/Isotype: Rabbit IgG