WB, IP, IF-IC, ChIP, C&R
H M R Mk
Endogenous
160, 180
Rabbit IgG
#Q96KQ7
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.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:100 |
| Immunofluorescence (Immunocytochemistry) | 1:3200 |
| Chromatin IP | 1:50 |
| CUT&RUN | 1:50 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human G9a 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).
- Tachibana, M. et al. (2001) J Biol Chem 276, 25309-17.
- Patnaik, D. et al. (2004) J Biol Chem 279, 53248-58.
- Tachibana, M. et al. (2002) Genes Dev 16, 1779-91.
- Tachibana, M. et al. (2005) Genes Dev 19, 815-26.
- Sampath, S.C. et al. (2007) Mol Cell 27, 596-608.
- Ueda, J. et al. (2006) J Biol Chem 281, 20120-8.
- Ogawa, H. et al. (2002) Science 296, 1132-6.
- Shi, Y. et al. (2003) Nature 422, 735-8.
- Nishio, H. and Walsh, M.J. (2004) Proc Natl Acad Sci USA 101, 11257-62.
- Estève, P.O. et al. (2006) Genes Dev 20, 3089-103.
- Tachibana, M. et al. (2007) EMBO J 26, 3346-59.
- 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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP C&R: CUT&RUN
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
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