| 货号 | 9718T |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human,Mouse,Rat,Monkey, |
| 来源宿主 | Rabbit IgG |
| 应用 | WB, IHC-P , F , IF-IC |
| 目标/特异性 | Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb detects endogenous levels of H2A.X only when phosphorylated at serine 139. |
| 使用方法 | WB(1:1000) IHC-P (1:480) F (1:200) IF-IC (1:200) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 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 |
| 参考文献 | 1 . Yuan, J. et al. (2010) FEBS Lett 584, 3717-24. 2 . Rogakou, E.P. et al. (1998) J Biol Chem 273, 5858-68. 3 . Burma, S. et al. (2001) J Biol Chem 276, 42462-7. 4 . Rogakou, E.P. et al. (1999) J Cell Biol 146, 905-16. 5 . Mukherjee, B. et al. (2006) DNA Repair (Amst) 5, 575-90. 6 . Solier, S. et al. (2009) Mol Cell Biol 29, 68-82. 7 . Lu, C. et al. (2006) Mol Cell 23, 121-32. 8 . Lu, C. et al. (2008) FEBS Lett 582, 2703-8. 9 . Cook, P.J. et al. (2009) Nature 458, 591-6. 10 . Xiao, A. et al. (2009) Nature 457, 57-62. |
Immunohistochemical analysis of paraffin-embedded HT-29 cells untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb. 使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb兔单抗，免疫组化分析HT-29细胞石蜡切片，细胞分为untreated (左图)或UV-treated (右图)。 | |
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). 使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb 兔单抗(绿色)，共聚焦免疫荧光分析HeLa细胞，细胞分为untreated (左图)或UV-treated (右图)。DY-554 phalloidin标记微丝蛋白(红色)。 | |
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb. 使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb兔单抗，免疫组化分析人源结肠癌石蜡切片。 | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb, showing nuclear localization. 使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb兔单抗，免疫组化分析人源肺癌石蜡切片，结果显示为细胞核定位。 | |
Western blot analysis of extracts from untreated or UV-treated 293 cells, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (upper) or Histone H2A.X Antibody #2595 (lower). 使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb兔单抗 (上图)或Histone H2A.X Antibody #2595 (下图)，免疫印迹(Western blot)分析untreated或UV-treated 293细胞系中Phospho-Histone H2A.X和Histone H2A.X的蛋白水平。 | |
Flow cytometric analysis of HeLa cells, untreated (blue) or UV-treated (green), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb compared to a nonspecific negative control antibody (red). 与一个非特异阴性control antibody (红色)比较，使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb进行流式细胞仪分析HeLa细胞，细胞分为untreated (蓝色)或UV-treated (绿色)。 | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb in the presence of control peptide (left) or Phospho-Histone H2A.X (Ser139) Blocking Peptide #1260 (right). 使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb兔单抗，左图是control peptide，右图是Phospho-Histone H2A.X (Ser139) Blocking Peptide #1260，免疫组化分析人源乳腺癌组织石蜡切片。 | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma untreated (left) or lambda-phosphatase-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb. 使用Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb兔单抗，免疫组化分析人源肺癌组织石蜡切片，左图是untreated，右图是lambda-phosphatase-treated。 |
