Chromatin Regulation / Acetylation
- Chromatin Regulation / Acetylation
PhosphoSitePlus®：
H3
- 推奨プロトコール
Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb |
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| CSTコード | 包装 | 希望納入価格 (円) |
国内在庫  2012年2月8日11時35分 現在 |
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| #9728S | 100 μL | 57,000 | ログインすると国内在庫状況がご確認いただけます。
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| #9728P | 40 μL for Custom Sampler Kit |  Custom Antibody Sampler Kitの構成品を選択できます。 5本以上を選択し、ページ右上のCartから製品確定書を発行してください。 尚、構成品の単品販売は致しておりません。 |
シグナル伝達研究応援キャンペーン プレゼント *Pサイズのみ
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9728 の推奨プロトコール 最適な結果を得るために：Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。 注：各製品に最適化されたプロトコールをリンクしています。 |
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| 用途 (希釈倍率) | |
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| ウェスタンブロッティング (1:1,000)、免疫蛍光細胞染色 (IF-IC) (1:6,400)、ChIP (1:50) |
| 種交差性 | |
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| ヒト、マウス、ラット、サル |
| 特異性・感度 | |
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| 内在性レベルのLys27 がジメチル化されたHistone H3 タンパク質を検出します。モノメチル化されたLys27 にはある程度の交差性が見られますが、トリメチル化された、またはメチル化されていないHistone H3 タンパク質とは交差しません。さらに、モノメチル化、ジメチル化、トリメチル化されたHistone H3 (Lys4、Lys9、Lys36) およびH4 タンパク質 (Lys20) とも交差しません。 |
| 検出タンパク質の分子量 | |
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| 17 kDa |
| 使用抗原 | |
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| Lys27 がジメチル化したHistone H3 タンパク質のN末端周辺領域 (合成ペプチド) |
| 抗体の由来 | |
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| ウサギ |
| 貯法 | |
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| -20℃ |
| 社内データ |
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Western Blotting
Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb.
IF-IC
Confocal immumunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb (green). Actin filaments have been labled with DY-554 phalloidin (red).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4x106 HeLa cells and either 10 μl of Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb 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 GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human AFM Intron 1 Primers #5098. 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.
ELISA-Peptide
Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated di-methyl histone H3 (Lys27) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the di-methyl histone H3 (Lys27) peptide competed away binding of the antibody.
| バックグラウンド |
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The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1 has shown that methylation is a reversible epigenetic marker (9).
- Peterson, C.L. and Laniel, M.A. (2004) Curr. Biol. 14, R546-R551.
- Kubicek, S. et al. (2006) Ernst Schering Res. Found Workshop, 1-27.
- Lin, W. and Dent, S.Y. (2006) Curr. Opin. Genet. Dev. 16, 137-142.
- Lee, D.Y. et al. (2005) Endocr. Rev. 26, 147-170.
- Daniel, J.A. et al. (2005) Cell Cycle 4, 919-926.
- Shi, X. et al. (2006) Nature 442, 96-99.
- Wysocka, J. et al. (2006) Nature 442, 86-90.
- Wysocka, J. et al. (2005) Cell 121, 859-872.
- Trojer, P. and Reinberg, D. (2006) Cell 125, 213-217.
| 使用例 | |
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本製品は試験研究用です。