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#8173 Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb

 
CSTコード 包装
希望納入価格 (円)
国内在庫 i
2017年12月11日15時55分 現在
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#8173S100 μL71,000
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#8173T20 μL   for Custom Sampler Kit
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Histone H3 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性17Rabbit-20℃
種交差性 (社内試験済)
交差する可能性がある種 i

社内試験はしていませんが、配列が100%相同であるため反応すると推定される種

ヒト、マウス、ラット、サル ハムスター、アフリカツメガエル、ゼブラフィッシュ、モルモット、ウマ
8173 の推奨プロトコール i

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。
推奨プロトコールはCST社内試験の徹底的なバリデーションに基づいて作成されておりますので、正確かつ再現性の高い結果が得られます。

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。

 

ウェスタンブロッティング (1:1000)免疫蛍光細胞染色 (IF-IC) (1:200)フローサイトメトリー (1:100)クロマチン免疫沈降 (1:100)クロマチン免疫沈降シークエンス (1:100)

CST推奨プロトコールに欠かせない関連製品

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、14、18、23、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
ヒトのHistone H3 タンパク質のアセチル化Lys27 周辺領域 (合成ペプチド)

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社内データ

※下記の社内データは、すべて8173 の推奨プロトコールで実験した結果です。

ELISA

ELISA

Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to precoated acetyl-histone H3 (Lys27) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the acetyl-histone H3 (Lys27) peptide competed away binding of the antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and C2C12 cells, untreated (-) or treated (+) with Trichostatin A (TSA) #9950 (1 μM, 18 hr), using Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb (upper) or Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A (TSA) #9950 (1 uM, 4 hr; right), using Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Trichostatin A (TSA) #9950 (1 uM, Overnight; green) using Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 50 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across GAPDH, a known target gene of H3K27Ac (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Acetyl-Histone H3 (Lys27) (D5E4) 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 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.


バックグラウンド

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 (1,2). Histone acetylation occurs mainly on the amino-terminal tail domains of histones H2A (Lys5), H2B (Lys5, 12, 15, and 20), H3 (Lys9, 14, 18, 23, 27, 36 and 56), and H4 (Lys5, 8, 12, and 16) and is important for the regulation of histone deposition, transcriptional activation, DNA replication, recombination, and DNA repair (1-3). Hyper-acetylation of the histone tails neutralizes the positive charge of these domains and is believed to weaken histone-DNA and nucleosome-nucleosome interactions, thereby destabilizing chromatin structure and increasing the accessibility of DNA to various DNA-binding proteins (4,5). In addition, acetylation of specific lysine residues creates docking sites for a protein module called the bromodomain, which binds to acetylated lysine residues (6). Many transcription and chromatin regulatory proteins contain bromodomains and may be recruited to gene promoters, in part, through binding of acetylated histone tails. Histone acetylation is mediated by histone acetyltransferases (HATs), such as CBP/p300, GCN5L2, PCAF, and Tip60, which are recruited to genes by DNA-bound protein factors to facilitate transcriptional activation (3). Deacetylation, which is mediated by histone deacetylases (HDAC and sirtuin proteins), reverses the effects of acetylation and generally facilitates transcriptional repression (7,8).

使用例
 
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XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb

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