Chromatin Regulation / Acetylation
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PhosphoSitePlus®：
| CSTコード | 包装 | 希望納入価格 (円) |
国内在庫  2012年2月7日15時25分 現在 |
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| #9727S | 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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| #9727L | 300 μL | 137,000 |
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9727 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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下記ステップについては、データシートの右側もあわせてご参照ください。
IHC-P: 抗体希釈液 / 抗原賦活化
| 用途 (希釈倍率) | |
|---|---|
| ウェスタンブロッティング (1:1,000)、免疫沈降 (1:50)、免疫組織染色 (パラフィン) (1:200)、免疫蛍光細胞染色 (IF-IC) (1:4,000)、ChIP (1:100) |
| 種交差性 | |
|---|---|
| ヒト、マウス、ラット、サル、(アフリカツメガエル、ゼブラフィッシュ) |
| 特異性・感度 | |
|---|---|
| 内在性レベルのLys4 がトリメチル化されたHistone H3 タンパク質を検出します。Lys4 がジメチル化されたHistone H3 タンパク質と若干交差しますが、Lys4が非メチル化およびモノメチル化されたHistone H3 タンパク質とは交差しません。また、Lys9、Lys27 およびLys36 がメチル化されたHistone H3 タンパク質およびLys20 がメチル化されたHistone H4 タンパク質とも交差しません。 |
| 検出タンパク質の分子量 | |
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| 17 kDa |
| 使用抗原 | |
|---|---|
| Lys4 がトリメチル化されたHistone H3 タンパク質のN末端領域 (合成ペプチド) |
| 抗体の由来 | |
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| ウサギ |
| 貯法 | |
|---|---|
| -20℃ |
| ※括弧付きの動物種は、配列が100%相同であるため反応すると推定されます。 |
| 社内データ |
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Western Blotting
Western blot analysis of lysates from HeLa, NIH/3T3, C6 and COS cells, using Tri-Methyl-Histone H3 (Lys4) antibody.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Tri-Methyl-Histone H3 (Lys4) Antibody.
IF-IC
Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Tri-Methyl-Histone H3 (Lys4) Antibody (green, left) compared to an isotype control (right). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
ELISA
Tri-Methyl-Histone H3 (Lys4) Antibody specificity was determined by peptide ELISA. Each graph depicts a titration of this antibody and the corresponding reactivity toward the non-methyl, mono-methyl, di-methyl and tri-methyl states of the indicated histone H3 or H4 lysine residue.
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Tri-Methyl-Histone H3 (Lys4) Antibody 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 RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
| 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.
| 使用文献 |
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- Song, N. et al. (2011) Acta Histochem Cytochem 44, 183-90. Applications: IHC-P (paraffin)
本製品は試験研究用です。