| 货号 | 7262C |
| 描述 | CSTs 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. |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | CSTs 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. |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | 4C |
| 参考文献 | Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358. Lazebnik, Y. A. et al. (1994) Nature 371, 346-347. Cohen, G.M. (1997) Biochem. J. 326, 1-16. Nicholson, D. W. et al. (1995) Nature 376, 37-43. Tewari, M. et al. (1995) Cell 81, 801-809. Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539. |
Figure 1: Treatment of HeLa cells with staurosporine stimulates cleavage of PARP at Asp214 as detected by PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit #7262. Absorbance at 450 nm is shown in the top figure, while the corresponding Western blot using Cleaved PARP (Asp214) Antibody #9548 (left panel) or PARP Antibody #9542 (right panel) is shown in the bottom figure. #9542 detects both full-length and cleaved PARP. 图1：使用PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit #7262检测 staurosporine（星形孢菌素）处理的HeLa细胞的裂解PARP（Asp214）蛋白的含量。450nm吸光值见顶图，免疫印迹检测到的裂解PARP（Asp214）蛋白水平见底图，使用抗体为Cleaved PARP (Asp214) Antibody #9548（左图）或PARP Antibody #9542（右图）。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and staurosporine (STO) treated HeLa cells and the absorbance at 450 nm is shown. HeLa cells (80% confluent) were treated with staurosporine (1 µM) for 3 hours. Absorbance at 450 nm is shown in the top figure, while the corresponding Western blot using Cleaved PARP Antibody #9548 is shown in the bottom figure. 图2：未处理和 staurosporine (STO)处理的Hela细胞裂解液蛋白浓度和试剂盒测得的450nm光密度读数之间的线性关系。Staurosporine（星形孢菌素）（1 uM）处理HeLa细胞（80％铺满）3小时。450nm吸光值见顶图，免疫印迹检测到的裂解PARP（Asp214）蛋白水平见底图，使用抗体为Cleaved PARP (Asp214) Antibody #9548。 |
