| 货号 | 5438S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F |
| 目标/特异性 | Phospho-Histone H2A.X (Ser139/Tyr142) Antibody recognizes endogenous levels of histone H2A.X protein only when phosphorylated at Ser139 or Tyr142, or dually phosphorylated on both residues. This antibody does not cross-react with other histone proteins. |
| 使用方法 | WB(1:1000) IP (1:50) F (1:200) IF-IC (1:100) |
| 供应商 | CST |
| 背景 | 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. |
| 存放说明 | -20C |
| 计算分子量 | 15 |
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). 使用Phospho-Histone H2A.X (Ser139/Tyr142) Antibody (绿色)，共聚焦免疫荧光分析untreated (左图)或UV-treated (右图)的COS-7细胞。DY-554 phalloidin标记微丝蛋白(红色)。 | |
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). 使用Phospho-Histone H2A.X (Ser139/Tyr142) Antibody (上图)或Histone H2A.X Antibody #2595 (下图)，免疫印迹(Western blot)分析untreated或UV-treated 293细胞。 | |
Flow cytometric analysis of HeLa cells, untreated (blue) or UV-treated (green), using Phospho-Histone H2A.X (Ser139/Tyr142) Antibody. 使用Phospho-Histone H2A.X (Ser139/Tyr142) Antibody标记，流式细胞仪分HeLa细胞，细胞分为untreated (蓝色)或UV-treated (绿色)。 |
