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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| #9649S | 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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| #9649P | 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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9649 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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- 9649:
- ChIP Agarose ChIP Magnetic Flow 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) | |
|---|---|
| ウェスタンブロッティング (1:1,000)、免疫組織染色 (パラフィン) (1:800)、免疫蛍光細胞染色 (IF-IC) (1:400)、ChIP (1:50)、フローサイトメトリー (1:200) |
| 種交差性 | |
|---|---|
| ヒト、マウス、ラット、サル、ゼブラフィッシュ |
| 特異性・感度 | |
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| 内在性レベルのLys9 がアセチル化されたHistone H3 タンパク質を検出します。他のアセチル化されたHistone タンパク質とは交差しません。 |
| 検出タンパク質の分子量 | |
|---|---|
| 17 kDa |
| 使用抗原 | |
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| Lys9 がアセチル化された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℃ |
| 社内データ |
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Western Blotting
Western blot analysis of lysates from HeLa and NIH/3T3 cells, untreated or TSA-treated (400 nM for 18 hours) using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human osteosarcoma using Acetyl-Histone H3 (Lys 9) (C5B11) Rabbit mAb.
Flow Cytometry
Flow cytometric analysis of untreated HeLa cells using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 versus propidium iodide (DNA content). Note positive staining in cycling cells (box).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with TSA #9950 (right), using Acetyl-Histone H3 (Lys9) (C5B11) 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 Acetyl-Histone H3 (Lys9) (C5B11) 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.
| バックグラウンド |
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Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).
- Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
- Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41.
- Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5.
- Cheung, P. et al. (2000) Cell 103, 263-71.
- Bernstein, B.E. and Schreiber, S.L. (2002) Chem Biol 9, 1167-73.
- Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
- Thorne, A.W. et al. (1990) Eur J Biochem 193, 701-13.
- Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
- Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
- Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.
- Dai, J. et al. (2005) Genes Dev 19, 472-88.
| 使用文献 |
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- Kong, D.K. et al. (2010) Mol Biol Cell 21, 1335-49. Applications: ChIP
- Zhao, J.X. et al. (2011) J Biol Chem 286, 16426-34. Applications: Western Blotting
本製品は試験研究用です。