| 货号 | 9947T |
| 描述 | This kit provides an economical means to analyze major signaling checkpoints in response to DNA damage. The kit contains primary and secondary antibodies to perform four Western blots with each antibody.该试剂盒为研究DNA损伤所致的主要信号检验点提供了一个经济的方式。该试剂盒提供了足够4次western blot实验的一抗和二抗。 |
| 目标/特异性 | All antibodies in the DNA Damage Antibody Sampler Kit recognize their targets proteins only when modified at the indicated site. |
| 供应商 | CST |
| 背景 | Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are PI3 Kinase-related kinase (PIKK) family members that phosphorylate multiple substrates on serine or threonine residues that are followed by a glutamine in response to DNA damage or replication blocks (1-3). p53 is phosphorylated by ATM, ATR and DNA-PK at Ser15. This phosphorylation impairs the ability of MDM2 to bind p53, promoting both the accumulation and activation of p53 in response to DNA damage (4,5). Chk1 and Chk2, downstream protein kinases of ATM/ATR, plays an important role in DNA damage checkpoint control, embryonic development and tumor suppression (6). Chk1 is phosphorylated at Ser280 and Ser296 following DNA damage. The amino-terminal domain of Chk2 contains a series of seven serine or threonine residues, including Thr68, each followed by glutamine (SQ or TQ motif). After DNA damage by ionizing radiation (IR), UV irradiation or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR (7-9). The breast cancer susceptibility proteins BRCA1 and BRCA2 are frequently mutated in cases of hereditary breast and ovarian cancers and have roles in multiple processes related to DNA damage, repair, cell cycle progression, transcription, ubiquitination and apoptosis. Numerous DNA-damage induced phosphorylation sites on BRCA1 have been identified, including serine 1524, and kinases activated in a cell cycle-dependent manner, including Aurora A and CDK2, can also phosphorylate BRCA1. IR, DNA and radiometric-induced DNA damage also results in rapid phosphorylation of the histone H2A family member H2A.X at Ser139 by ATM (10,11). Within minutes following DNA damage, Ser139-phosphorylated H2A.X localizes to sites of DNA damage at subnuclear foci (12).共济失调毛细血管扩张症突变蛋白激酶(ATM)和共济失调毛细血管扩张症以及Rad3相关激酶(ATR)属于PI3激酶相关激酶(PIKK)家族成员，能够磷酸化DNA损伤或者复制阻遏中跟随谷氨酰胺的多重底物的丝氨酸或者苏氨酸残基(1-3)。 ATM，ATR和DNA-PK 能够磷酸化p53的 15位丝氨酸。这种磷酸化削弱了MDM2蛋白结合p53 的能力，促进两者的积累和p53的激活以应答DNA损伤(4,5)。Chk1和Chk2，是ATM/ATR蛋白激酶的下游，在DNA损伤检验点控制、胚胎发育和肿瘤抑制中起着重要的作用(6)。Chk1 Ser280和Ser296的磷酸化紧随DNA损伤。Chk2的氨基末端结构域包含一个7丝氨酸或苏氨酸残基系列，包括Thr68，每个紧接着谷氨酰胺(SQ或 TQ基序)。电离辐射（IR），紫外光照射或羟基脲处理引起DNA损伤后，该区域的Thr68和其它位点被ATM/ ATR磷酸化(7-9)。乳腺癌易感蛋白BRCA1和BRCA2在遗传性乳腺癌和卵巢癌的情况下频繁突变，在多个相关的进程中起作用，如DNA损伤、修复、细胞周期进程、转录、泛素化和细胞凋亡。大量的DNA损伤诱导的BRCA1磷酸化位点已经确定，包括1524位丝氨酸，同时细胞周期依赖方式激活的激酶，包括极光激酶A和CDK2，也可以磷酸化BRCA1。IR，DNA和辐射诱导的DNA损伤也可导致组蛋白H2A家族成员H2A.X的Ser139被ATM迅速磷酸化(10,11)。DNA损伤后的几分钟内， Ser139磷酸化的H2A.X定位到亚核灶的DNA损伤位点(12)。Application References
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| 存放说明 | -20C |
| 参考文献 | 1 . Kastan, M.B. and Lim, D.S. (2000) Nat Rev Mol Cell Biol 1, 179-86. 2 . Abraham, R.T. (2004) DNA Repair (Amst) 3, 883-7. 3 . Rogakou, E.P. et al. (1998) J Biol Chem 273, 5858-68. 4 . Shechter, D. et al. (2004) DNA Repair (Amst) 3, 901-8. 5 . Burma, S. et al. (2001) J Biol Chem 276, 42462-7. 6 . Shieh, S.Y. et al. (1997) Cell 91, 325-34. 7 . Rogakou, E.P. et al. (1999) J Cell Biol 146, 905-16. 8 . Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7. 9 . Martinho, R.G. et al. (1998) EMBO J. 17, 7239-17249. 10 . Matsuoka, S. et al. (2000) Proc Natl Acad Sci USA 97, 10389-94. 11 . Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762-765. 12 . Ahn, J.Y. et al. (2000) Cancer Res. 60, 5934-5936. |
Western blot analysis of extracts from HeLa, COS, NIH/3T3 and C6 cells, untreated or UV-treated, using Phospho-Chk1 (Ser345) (133D30) Rabbit mAb #2348. | |
Western blot analysis of extracts from 293 cells, untreated or UV-treated (100 mJ, 4 hr recovery), using Phospho-ATM (Ser1981) (D6H9) Rabbit mAb #5883 (upper) or ATM (D2E2) Rabbit mAb #2873 (lower).Western blot 方法检测未处理或紫外处理的(100 mJ, 恢复4小时)的293细胞提取物，使用的抗体为Phospho-ATM (Ser1981) (D6H9) Rabbit mAb #5883 (上图) 或 ATM (D2E2) Rabbit mAb #2873 (下图)。 | |
Western blot analysis of extracts from 293 cells, untreated or UV-treated (100 mJ, 4 hr recovery), using Phospho-ATM (Ser1981) (D6H9) Rabbit mAb (upper) or ATM (D2E2) Rabbit mAb #2873 (lower). | |
Western blot analysis of extracts from untreated or UV-treated 293 cells, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb # 9718 (upper) or Histone H2A.X Antibody #2595 (lower).Western blot 方法检测未处理或紫外处理的293细胞提取物，使用的抗体为 Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb # 9718 (上图) 或 Histone H2A.X Antibody #2595 (下图)。 | |
Flow cytometric analysis of HeLa cells, untreated (blue) and UV-treated (green), using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb. | |
Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or UV-treated (right), using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb. | |
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). | |
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded HT-29 cell pellets, control (left) or UV-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb. | |
Western blot analysis of extracts from HeLa cells, untreated or UV-treated, using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb. | |
Immunoprecipitation of phospho-chk2 from UV-treated HT29 cells using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb followed by western blot using the same antibody. | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb. | |
Flow cytometric analysis of untreated Jurkat cells, using Phospho-Chk2 (Thr68) (C13C1) Rb mAb versus propidium iodide (DNA content). The boxed population indicates phospho-Chk2 (Thr68)-positive cells. | |
Western blot analysis of extracts from 293 cells, untreated, UV-treated or doxorubicin-treated (0.5 µM), using Phospho-Chk2 (Thr68) Antibody #2661.Western blot方法检测未处理、紫外处理或阿霉素(0.5 µM)处理的293细胞，使用的抗体为Phospho-Chk2 (Thr68) Antibody 。 |
