Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

G9a/EHMT2 (C6H3) Rabbit mAb #3306

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC H M R Mk (B) (Pg) (Hr) Endogenous 140, 165 Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Pg=Pig  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

G9a/EHMT2 (C6H3) Rabbit mAb 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.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human G9a/EHMT2 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and 293 cells using G9a/EHMT2 (C6H3) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using G9a/EHMT2 (C6H3) Rabbit mAb (green). Actin filaments have been labeled with DY-555 phalloidin (red).

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 in vitro (1,2). However, in vivo 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).

  1. Tachibana, M. et al. (2001) J Biol Chem 276, 25309-17.
  2. Patnaik, D. et al. (2004) J Biol Chem 279, 53248-58.
  3. Tachibana, M. et al. (2002) Genes Dev 16, 1779-91.
  4. Tachibana, M. et al. (2005) Genes Dev 19, 815-26.
  5. Sampath, S.C. et al. (2007) Mol Cell 27, 596-608.
  6. Ueda, J. et al. (2006) J Biol Chem 281, 20120-8.
  7. Ogawa, H. et al. (2002) Science 296, 1132-6.
  8. Shi, Y. et al. (2003) Nature 422, 735-8.
  9. Nishio, H. and Walsh, M.J. (2004) Proc Natl Acad Sci USA 101, 11257-62.
  10. Estève, P.O. et al. (2006) Genes Dev 20, 3089-103.
  11. Tachibana, M. et al. (2007) EMBO J 26, 3346-59.
  12. Kondo, Y. et al. (2007) Hepatol Res 37, 974-83.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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