Revision 3

#3306Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IF-IC, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

160,180

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q96KQ7

Entrez-Gene Id:

10919

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
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.

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.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Bovine, Pig, Horse

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 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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 3
#3306

G9a/EHMT2 (C6H3) Rabbit mAb

Western Blotting Image 1: G9a/EHMT2 (C6H3) Rabbit mAb Expand Image
使用 G9a/EHMT2 (C6H3) Rabbit mAb 对 HeLa 和 293 细胞系提取物进行蛋白质印迹分析。
Immunofluorescence Image 1: G9a/EHMT2 (C6H3) Rabbit mAb Expand Image
使用 G9a/EHMT2 (C6H3) Rabbit mAb(绿色)对 Hela 细胞进行共聚焦免疫荧光分析。肌动蛋白丝可用 DY-555 Phalloidin(红色)标记。
Chromatin Immunoprecipitation Image 1: G9a/EHMT2 (C6H3) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 K562 细胞中提取的交联染色质,在加入 G9a/EHMT2 (C6H3) 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀法分析。使用 SimpleChIP® Human ZNF19 Intron 3 Primers #98428、人 VRK3 intron 11 primers 和 SimpleChIP® Human GAPDH Exon 1 Primers #5516,通过实时 PCR 对富集的 DNA 进行量化。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。