PathScan? Cleaved PARP (Asp214) Sandwich ELISA Kit

• 产品编号：CST-7262C      品牌：CST       原厂货号：7262C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

CST的PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit能够检测内源的裂解的PARP (Asp214)蛋白。如图1所示，使用Cleaved PARP (Asp214) ELISA Kit #7262检测到 staurosporine 处理的HeLa细胞中裂解的PARP (Asp214)蛋白显著减少。经内部测试验证，该试剂盒能够检测指定物种的蛋白质，同时也可以检测其他物种的同源蛋白。CST的PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit采用了固相夹心酶联免疫吸附法（ELISA）检测内源的裂解的PARP (Asp214)蛋白。微孔中包被了cleaved PARP (Asp214) Mouse mAb 鼠单抗。细胞裂解液孵育后，裂解的PARP (Asp214)蛋白被包被的抗体捕获。充分洗涤后，添加PARP Rabbit mAb 兔单抗检测捕获的裂解的PARP蛋白。再用Anti-rabbit IgG, HRP-linked Antibody #7074识别结合的抗体。添加HRP底物，TMB 显色。测得的吸光值与裂解的PARP (Asp214)蛋白的量成正比。试剂盒中的抗体为该试剂盒所特制。PARP，是核多聚(ADP-核糖) 聚合酶，分子量116kDa，参与环境胁迫应答中的DNA修复(1)。在体外该蛋白可被许多ICE样 caspases裂解(2,3)，在体内是caspase-3裂解的主要靶蛋白之一(4,5)。在人体中PARP在位点Asp214 和 Gly215被裂解，裂解后PARR的N末端DNA 结合结构域(24 kDa)与C端催化结构域(89 kDa) 分离(2,4)。PARP协助细胞维持生存能力；PARP的裂解促进细胞崩解并可以作为细胞凋亡的标记物(6）。

1. Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358.
2. Lazebnik, Y. A. et al. (1994) Nature 371, 346-347.
3. Cohen, G.M. (1997) Biochem. J. 326, 1-16.
4. Nicholson, D. W. et al. (1995) Nature 376, 37-43.
5. Tewari, M. et al. (1995) Cell 81, 801-809.
6. Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit detects endogenous levels of cleaved PARP (Asp214) protein. As shown in Figure 1, using Cleaved PARP (Asp214) ELISA Kit #7262, a significant induction of cleaved PARP (Asp214) is detected in HeLa cells treated with staurosporine. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST's PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of cleaved PARP (Asp214) protein. A cleaved PARP (Asp214) Mouse mAb* has been coated onto the microwells. After incubation with cell lysates, cleaved PARP (Asp214) protein is captured by the coated antibody. Following extensive washing, PARP Rabbit mAb* is added to detect the captured cleaved PARP protein. Anti-rabbit IgG, HRP-linked Antibody #7074* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of cleaved PARP (Asp214) protein. 
 * Antibodies in kit are custom formulations specific to kit.

Background

PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (1). This protein can be cleaved by many ICE-like caspases in vitro (2,3) and is one of the main cleavage targets of caspase-3 in vivo (4,5). In human PARP, the cleavage occurs between Asp214 and Gly215, which separates the PARP amino-terminal DNA binding domain (24 kDa) from the carboxy-terminal catalytic domain (89 kDa) (2,4). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (6).

1. Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358.
2. Lazebnik, Y. A. et al. (1994) Nature 371, 346-347.
3. Cohen, G.M. (1997) Biochem. J. 326, 1-16.
4. Nicholson, D. W. et al. (1995) Nature 376, 37-43.
5. Tewari, M. et al. (1995) Cell 81, 801-809.
6. Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).