Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

G9a/EHMT2 (C6H3) Rabbit mAb #3306

EHMT2   G9a   histone-methyltransferase   HMT  

No. Size Price
3306S 100 µl ( 10 western blots ) ¥3,250.00 现货查询 购买询价 防伪查询
3306T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价 防伪查询
3306 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 140, 165 Rabbit IgG
IF-IC 1:50
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,


Species predicted to react based on 100% sequence homology: Bovine, Pig, Horse,

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.

G9a/EHMT2 (C6H3) Rabbit mAb兔单抗能够检测内源性G9a/EHMT2总蛋白(165 kDa G9a-L和140 kDa G9a-S亚型)。该抗体不能与其它组蛋白甲基转移酶包括GLP发生交叉反应。

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.

使用G9a/EHMT2 (C6H3) Rabbit mAb兔单抗,免疫印迹(Western blot)分析HeLa和293细胞系中G9a/EHMT2的蛋白水平。



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

使用G9a/EHMT2 (C6H3) Rabbit mAb 兔单抗(绿色)标记,共聚焦免疫荧光分析HeLa细胞。DY-554 phalloidin标记微丝蛋白(红色)。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 K562 cells and either 10 μl of G9a/EHMT2 (C6H3) or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ZNF19 Intron 3 Primers #98428, human VRK3 intron 11 primers, and SimpleChIP® Human GAPDH Exon 1 Primers #5516. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.


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).

G9a也称为Euchromatic histone-lysine N-methyltransferase 2 (EHMT2),它是一个组蛋白赖氨酸甲基转移酶家族成员,该家族每个成员都含有一个保守的起催化作用的SET结构域,该结构起初在果蝇Su[var]3-9、zeste增强子和Trithorax蛋白中被鉴定(1)。在体外,重组的G9a能够使histone H3在Lys9和Lys27位点发生单甲基、双甲基和三甲基化作用(1,2)。然而,在体内G9a形成一个GLP的复合物,它是一个G9a相关的组蛋白甲基转移酶,并且它与这些蛋白一起在主要的常染色质的histone H3的Lys9位点发挥单核双甲基转移酶作用,这能产生转录抑制标记物,其能有助于基因沉默(3,4)。G9a能使自身在Lys165位点发生甲基化,该修饰能调节HP1抑制蛋白与G9a/GLP复合物的关联(5)。G9a/GLP 复合物也包含Wiz蛋白,Wiz蛋白是一个锌指蛋白,它对于G9a/GLP异源二聚体和复合物稳定性是需要的(6)。Wiz蛋白包含两个CtBP共抑制结合位点,这能够调节G9a/GLP复合物与CtBP共抑制复合物的关联(6)。此外,G9a和GLP是其它大转录共抑制复合物的成分,例如涉及E2F6和CDP/cut(7-9)。G9a与DNMT1相互作用,并且这两个蛋白对于DNA甲基化和在复制点的histone H3 (Lys9)是需要的,在DNA复制期间,这提供了一个在histone H3 Lys9和CpG甲基化之间功能性联系(10)。G9a活性对于减数分裂前期是关键的,因为突变老鼠生殖细胞系G9a缺陷显示一个大量的成熟配子减少(11)。因此,G9a有助于在缺氧应激下 di-methyl histone H3 (Lys9)的整体水平增加,以及增加的G9a表达水平与肝癌有关联(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.

U.S. Patent No. 5,675,063.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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