DNA Damage Antibody Sampler Kit

• 产品编号：CST-9947S      品牌：Cell Signaling Technology       原厂货号：9947S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

DNA Damage Antibody Sampler Kit 中的所有抗体都能够识别其在特定位点修饰了的靶蛋白。多克隆抗体是由合成的人源的针对ATR蛋白丝氨酸（428位）、BRCA1蛋白丝氨酸（1524位）、Chk1蛋白丝氨酸（296位）、Chk2蛋白苏氨酸（68位）的磷酸化肽段免疫动物，采用蛋白A和多肽亲和层析技术生产的。单克隆抗体是由合成的人源的针对Histone H2A.X蛋白丝氨酸（139位）、ATM蛋白丝氨酸（1981位）、p53蛋白丝氨酸（15位）的磷酸化肽段免疫动物生产的。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。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)。

原厂资料：

Specificity / Sensitivity

All antibodies in the DNA Damage Antibody Sampler Kit recognize their targets proteins only when modified at the indicated site.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser428 of human ATR; Ser1524 of human BRCA1; Ser296 of human Chk1; or Thr68 of human Chk2. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding, Ser139 of Histone H2A.X; Ser1981 of human ATM; or Ser15 of human p53

Description

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.

Background

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).

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not
aliquot the antibodies.