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#50929 PathScan® Phospho-Histone H2A.X (Ser139) Sandwich ELISA Kit

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2019年3月22日15時35分 現在
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#50929C1 Kit105,000

Histone H2A.X 製品一覧

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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。


キット内容 容量 液色
Histone H2A.X Rabbit mAb Coated Microwells 96 tests
P-Histone H2A.X (Ser139) Mouse Detection mAb 1 ea Green (Lyophilized)
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
TMB Substrate #7004 11 ml Colorless
STOP Solution #7002 11 ml Colorless
Sealing Tape 2 ea
ELISA Wash Buffer (20X) #9801 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) #9803 15 ml Yellowish

Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Kit 情報
内在性レベルのSer139 がリン酸化されたHistone H2A.X タンパク質を検出します。
4℃ (Lysis Buffer は-20℃)
※Lysis Buffer は保存温度が-20℃となっておりますが、輸送時においては保冷 (冷媒梱包) とさせて頂いています。本品受領後は速やかに冷凍庫 (-20℃) での保存をお願いいたします。

ELISA - Western correlation

ELISA - Western correlation

Figure 1: Phospho-Histone H2A.X (Ser139) protein from untreated (-) and UV-treated (+) 293 cells is detected by PathScan® Phospho-Histone H2A.X (Ser139) Sandwich ELISA kit. A lower signal of phospho-Histone H2A.X protein is seen in the untreated lysate. However, similar levels of total Histone H2A.X protein from both untreated and treated lysates are shown by western analysis. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb #9718 is shown in the bottom figure.



Figure 2: The relationship between protein concentration of lysates from untreated and UV-treated 293 cells and the absorbance at 450 nm as detected by the PathScan® Phospho-Histone H2A.X Ser139 Sandwich ELISA Kit is shown.

Unstarved 293 cells (80-90% confluence) were treated with 100 mJ/cm2 UV with 1 hr recovery at 37ºC, and then lysed with Cell Lysis Buffer (#9803).


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.

2577   Phospho-Histone H2A.X (Ser139) Antibody
2595   Histone H2A.X Antibody
7076   Anti-mouse IgG, HRP-linked Antibody
7631   Histone H2A.X (D17A3) XP® Rabbit mAb
9718   Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb
9803   Cell Lysis Buffer (10X)
9808   Phosphate Buffered Saline (PBS-20X)
9809   Phosphate Buffered Saline with Tween® 20 (PBST-20X)

PathScan is a trademark of Cell Signaling Technology, Inc.
FastScan™ ELISA is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.


PathScan® Phospho-Histone H2A.X (Ser139) Sandwich ELISA Kit

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