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#5438 Phospho-Histone H2A.X (Ser139/Tyr142) Antibody

 
CSTコード 包装
希望納入価格 (円)
国内在庫 i
2019年8月20日15時55分 現在
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#5438S100 μL66,000
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感度分子量 (kDa)抗体の由来貯法
内在性15Rabbit-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 -
5438 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)免疫沈降 (1:50)免疫蛍光細胞染色 (IF-IC) (1:100)フローサイトメトリー (1:200)

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Ser139 とTyr142 の片方、もしくは両方がリン酸化されたHistone H2A.X タンパク質を検出します。他のHistone タンパク質とは交差しません。
使用抗原
ヒトのHistone H2A.X タンパク質のSer139 とTyr142 周辺領域 (合成リン酸化ペプチド)

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社内データ

※下記の社内データは、すべて5438 の推奨プロトコールで実験した結果です。

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated, using Phospho-Histone H2A.X (Ser139/Tyr142) Antibody (upper) or Histone H2A.X Antibody #2595 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS-7 cells, untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139/Tyr142) Antibody (green). Actin filaments were labeled with DY-554 phalloidin (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or UV-treated (green), using Phospho-Histone H2A.X (Ser139/Tyr142) Antibody.


バックグラウンド

Histone H2A.X is a variant histone that represents approximately 10% of the total H2A histone proteins in normal human fibroblasts (1). H2A.X is required for checkpoint-mediated cell cycle arrest and DNA repair following double-stranded DNA breaks (1). DNA damage, caused by ionizing radiation, UV-light, or radiomimetic agents, results in rapid phosphorylation of H2A.X at Ser139 by PI3K-like kinases, including ATM, ATR, and DNA-PK (2,3). Within minutes following DNA damage, H2A.X is phosphorylated at Ser139 at sites of DNA damage (4). This very early event in the DNA-damage response is required for recruitment of a multitude of DNA-damage response proteins, including MDC1, NBS1, RAD50, MRE11, 53BP1, and BRCA1 (1). In addition to its role in DNA-damage repair, H2A.X is required for DNA fragmentation during apoptosis and is phosphorylated by various kinases in response to apoptotic signals. H2A.X is phosphorylated at Ser139 by DNA-PK in response to cell death receptor activation, c-Jun N-terminal Kinase (JNK1) in response to UV-A irradiation, and p38 MAPK in response to serum starvation (5-8). H2A.X is constitutively phosphorylated on Tyr142 in undamaged cells by WSTF (Williams-Beuren syndrome transcription factor) (9,10). Upon DNA damage, and concurrent with phosphorylation of Ser139, Tyr142 is dephosphorylated at sites of DNA damage by recruited EYA1 and EYA3 phosphatases (9). While phosphorylation at Ser139 facilitates the recruitment of DNA repair proteins and apoptotic proteins to sites of DNA damage, phosphorylation at Tyr142 appears to determine which set of proteins are recruited. Phosphorylation of H2A.X at Tyr142 inhibits the recruitment of DNA repair proteins and promotes binding of pro-apoptotic factors such as JNK1 (9). Mouse embryonic fibroblasts expressing only mutant H2A.X Y142F, which favors recruitment of DNA repair proteins over apoptotic proteins, show a reduced apoptotic response to ionizing radiation (9). Thus, it appears that the balance of H2A.X Tyr142 phosphorylation and dephosphorylation provides a switch mechanism to determine cell fate after DNA damage.

使用例
 
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9999   Nonfat Dry Milk

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

Phospho-Histone H2A.X (Ser139/Tyr142) Antibody

Immune Cell Signaling Pathways

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