| 货号 | 8342T |
| 描述 | The UV Induced DNA Damage Response Antibody Sampler Kit offers an economical means of investigating proteins involved in the cellular response to UV-induced DNA damage. The kit contains enough primary and secondary antibody to perform four western blot experiments per primary.UV Induced DNA Damage Response Antibody Sampler Kit能够经济地研究紫外诱导的DNA损伤引起的细胞反应中的相关蛋白。该试剂盒内含足够4次western blot实验的一抗和二抗。 |
| 目标/特异性 | Antibodies detect endogenous levels of the respective target proteins. |
| 供应商 | CST |
| 背景 | Exposure to ultraviolet radiation (UV) has a profound impact on human health and disease (1). Low level UV exposure induces the production of vitamin D and is a key regulator of calcium metabolism. Conversely, overexposure to UV is associated with an increased risk of cancer, immunosuppression, and many eye disorders, such as cataracts. Photons of UV light can directly damage DNA causing thymine dimers and other pyrimidine dimers between adjacent bases (2). Free radicals and reactive oxygen species induced by UV exposure also result in DNA lesions and have been linked to malignant melanoma (3). DNA damage from replicative stress and genotoxic agents like UV activate the ATR-mediated checkpoint pathway and stimulate DNA repair, cell cycle arrest, and apoptosis (4). ATR recruitment to sites of DNA damage and activation depends, at least in part, on interaction with the complex of single-stranded DNA, Replication Protein A (RPA), and direct binding to the ATR-associated adapter protein, ATRIP (5). In addition, the Rad17-RFC and Rad9-Rad1-Hus1 (9-1-1) protein complexes are independently recruited with TopBP1 to fully activate the checkpoint response (6,7). BRIT1 (MCPH1) is required for UV-induced formation of ATR, RPA, and p-Rad17 foci at sites of DNA damage (8-10) and may regulate the expression of several DNA damage response proteins (11). Once activated, ATR phosphorylates a number of mediators, including histone H2AX Ser139 and Chk1 kinase at Ser345. H2AX phosphorylation is a marker of DNA damage. Complete loss of H2AX results in reduced Chk1 activation and impaired survival of cells after UV exposure (12). Chk1 and Chk2 kinase activation is essential for checkpoint-mediated control of cell cycle progression (4). Checkpoint kinases stimulate cell cycle arrest by phosphorylation of a group of tyrosine phosphatases known as Cdc25A, Cdc25B, and Cdc25C (13 -15). Both Chk1 and Chk2 kinases phosphorylate Cdc25C at Ser216 in response to DNA damage and stimulate arrest (16-17).暴露于紫外线(UV)对人类健康和疾病具有深远的影响(1)。低水平的紫外线照射能够诱导维生素D的产生且是钙代谢的关键调节子。相反，过度暴露于紫外线与癌症、免疫抑制以及许多眼部疾病如白内障的风险增加有关。紫外光的光子可直接损伤DNA造成胸腺嘧啶二聚体和相邻碱基之间的其他嘧啶二聚体(2)。紫外线照射引起的自由基和活性氧可导致DNA损伤并与恶性黑色素瘤有关(3)。来自复制应激和遗传毒性药物如紫外线的DNA损伤能够激活ATR-介导的检验点途径并促进DNA修复、细胞周期阻滞和细胞凋亡(4)。ATR招募至DNA损伤点和激活至少部分取决于和单链DNA复制蛋白A(RPA)相互作用以及直接结合ATR-相关的接头蛋白ATRIP(5)。此外，Rad17-RFC 和Rad9-Rad1-Hus1 (9-1-1)蛋白质复合物被独立地招募到TopBP1从而完全激活检验点反应(6,7)。BRIT1 (MCPH1)为紫外线引起的DNA损伤的位点形成ATR，RPA，和p-Rad17所必需(8-10)，并可以调节几个DNA损伤反应蛋白的表达(11)。一旦被激活，ATR能够磷酸化一些中间体，包括组蛋白H2AX Ser139和Chk1激酶Ser345。H2AX磷酸化是DNA损伤的一个标志物。H2AX的完全缺失能够导致Chk1激活的极少并削弱紫外暴露后细胞的存活(12)。Chk1和Chk2激酶活化对于检验点介导的细胞周期进程控制是必不可少的(4)。检验点激酶通过磷酸化一组酪氨酸磷酸酶Cdc25A、Cdc25B和Cdc25C来刺激细胞周期阻滞(13 -15)。Chk1和Chk2激酶都能够磷酸化Cdc25C蛋白的216位丝氨酸来应答DNA损伤反应和刺激捕获(16-17)。Application References |
| 存放说明 | -20C |
| 参考文献 | 1 . Peng G et al. (2009) Nat Cell Biol 11, 865–72 2 . von Thaler, A.K. et al. (2010) Exp Dermatol 19, 81-8. 3 . Rastogi, R.P. et al. (2010) J Nucleic Acids 2010, 592980. 4 . Narayanan, D.L. et al. (2010) Int J Dermatol 49, 978-86. 5 . Zou, L. and Elledge, S.J. (2003) Science 300, 1542-8. 6 . Zhou, B.B. and Elledge, S.J. (2000) Nature 408, 433-439. 7 . Blasina, A. et al. (1999) Curr. Biol. 9, 1-10. 8 . Furnari, B. et al. (1999) Mol. Biol. Cell 10, 833-845. 9 . Zou, L. et al. (2002) Genes Dev 16, 198-208. 10 . Mordes, D.A. and Cortez, D. (2008) Cell Cycle 7, 2809-12. 11 . Rai, R. et al. (2006) Cancer Cell 10, 145-57. 12 . Lin, S.Y. et al. (2010) Yonsei Med J 51, 295-301. 13 . Lin, S.Y. et al. (2005) Proc Natl Acad Sci U S A 102, 15105-9. 14 . Revet, I. et al. (2011) Proc Natl Acad Sci U S A 108, 8663-7. 15 . Mailand, N. et al. (2000) Science 288, 1425-9. 16 . Sanchez, Y. et al. (1997) Science 277, 1497-501. 17 . Matsuoka, S. et al. (1998) Science 282, 1893-7. |
Western blot analysis of extracts from various cell lines using Microcephalin-1/BRIT1 (D38G5) Rabbit mAb #4120.Western blot 方法检测不同细胞提取物，使用的抗体为 Microcephalin-1/BRIT1 (D38G5) Rabbit mAb #4120。 | |
Western blot analysis of extracts from Jurkat and HEL cells using ATRIP Antibody #2737.Western blot 方法检测Jurkat 和HEL 细胞提取物，使用的抗体为 ATRIP Antibody #2737. | |
Western blot analysis of Raw 264.7, SV-T2, and HT-29 cells, untreated or UV-treated (50 mJ, 30 min), using Phospho-ATR (Ser428) Antibody #2853. λ phosphatase (10,000 Units/ml, 1hr) was used to demonstrate the phospho-specificity of the antibody.Western blot 方法检测未处理和紫外处理(50 mJ)30分钟的Raw 264.7, SV-T2和HT-29细胞，使用的抗体为Phospho-ATR (Ser428) Antibody #2853。 λ 磷酸酶(10,000 Units/ml)处理1小时用以检测抗体的磷酸化特异性。 | |
Western blot analysis of extracts from various cell lines using RPA32 (4E4) Rat mAb #2208.Western blot 方法检测不同细胞提取物，使用的抗体为 RPA32 (4E4) Rat mAb #2208。 | |
Western blot analysis of extracts from various cell lines, untreated or UV-treated, using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #2348.Western blot方法检测未处理和紫外处理的各种细胞提取物，使用的抗体为Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #2348. | |
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). | |
Western blot analysis of extracts from various cell types using Microcephalin-1/BRIT1 (D38G5) Rabbit mAb. | |
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方法检测未处理和紫外处理的293细胞，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). | |
Western blot analysis of HT-29 cells, untreated, nocodazole-treated, and λ phosphotase-treated, using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb #4901 (upper) or cdc25C (5H9) Rabbit mAb #4688 (lower).Western blot方法检测未处理、诺考达唑处理和λ磷酸酶处理的HT-29细胞提取物，使用的抗体为 Phospho-cdc25C (Ser216) (63F9) Rabbit mAb #4901 (上图)或cdc25C (5H9) Rabbit mAb #4688 (下图). | |
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). | |
Western Blot analysis of HT29 cells, untreated, nocodazole-treated, and lambda phosphotase-treated, using Phospho-cdc25C (Ser216) antibody (upper), and cdc25C (5H9) Rabbit Mab, #4688, (lower). | |
Western blot analysis of extracts from various cell lines, using RPA32 (4E4) Rat mAb. | |
Western blot analysis of extracts from Jurkat and HEL cells using ATRIP Antibody. | |
Confocal immunofluorescent analysis of HeLa cells using ATRIP Antibody (green). Actin filaments have been labeled with Alexa Fluor®555 phalloidin (red). |
