Chromatin Regulation / Acetylation
- Chromatin Regulation / Acetylation
PhosphoSitePlus®：
H3
- 推奨プロトコール
Di-Methyl-Histone H3 (Lys4) (C64G9) 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17時20分 現在 |
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| #9725S | 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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| #9725P | 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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9725 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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- 9725:
- ChIP Agarose ChIP Magnetic IHC / Paraffin Immunofluorescence Immunoprecipitation Western Blotting
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） | |
|---|---|
| ウエスタンブロッティング（1:1,000）、免疫沈降（1:25）、免疫組織染色（パラフィン）（1:1,500）、免疫蛍光細胞染色（IF-IC）（1:800）、ChIP（1:50） |
| 種交差性 | |
|---|---|
| ヒト、マウス、ラット、サル、（全ての種） |
| 特異性・感度 | |
|---|---|
| 内在性レベルのLy4 がジメチル化されたHistone H3 タンパク質を検出します。Ly4 がメチル化されていない、モノメチル化、およびトリメチル化されたHistone H3 タンパク質とは交差しません。また、Lys9、Lys27、Lys36 がジメチル化、またはトリメチル化されたHistone H3 タンパク質とLys20がジメチル化、またはトリメチル化された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 | |
|---|---|
| 17 kDa |
| 使用抗原 | |
|---|---|
| Lys4 がジメチル化されたHistone H3 タンパク質のN末端領域（合成ペプチド） |
| 抗体の由来 | |
|---|---|
| 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 whole cell lysates from HeLa, NIH/3T3, C6 and COS cells using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human stomach carcinoma using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb.
IF-IC
Confocal immunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Di-Methyl-Histone H3 (Lys4) (C64G9) 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 RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, 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.
ELISA-Peptide
Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated di-methyl histone H3 (Lys4) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the di-methyl histone H3 (Lys4) 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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| 製品をご使用いただいて研究を発表されましたら、ぜひお知らせください。 |
本製品は試験研究用です。