Chromatin Regulation / Acetylation
Acetyl-Histone H3 (Lys27) Antibody
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| CSTコード |
包装 |
希望納入価格 (円) |
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| #4353S | 100 μL | 57,000 | |
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4353 の推奨プロトコール
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4353:
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ChIP Agarose
ChIP Magnetic
Immunoprecipitation
Western Blotting
| 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 (1:25)、ChIP (1:25) |
| 種交差性 | |
| ヒト、マウス、ラット、サル、(ニワトリ、ハムスター、ウシ、キイロショウジョウバエ、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Lys27 がアセチル化されたHistone H3 タンパク質を検出します。Lys9 がアセチル化されたHistone H3 タンパク質とわずかに交差しますが、Lys14、18、56 がアセチル化されたHistone H3 タンパク質とは交差しません。 |
| 使用抗原 | |
| Lys27 がアセチル化されたHistone H3 タンパク質のN末端周辺領域 (合成ペプチド) |
| ※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。 |
Western Blotting
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated with Trichostatin A #9950 (400 nM for 18 h), using Acetyl-Histone H3 (Lys27) Antibody (upper) and Histone H3 Antibody #9715 (lower).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 μl of Acetyl-Histone H3 (Lys27) 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 AFM Intron 1 Primers #5098, 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.
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).
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Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
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Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41.
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Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5.
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Cheung, P. et al. (2000) Cell 103, 263-71.
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Bernstein, B.E. and Schreiber, S.L. (2002) Chem Biol 9, 1167-73.
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Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
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Thorne, A.W. et al. (1990) Eur J Biochem 193, 701-13.
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Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
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Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
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Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.
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Dai, J. et al. (2005) Genes Dev 19, 472-88.
PhosphoSitePlus®：
H3