| 货号 | 7190C |
| 描述 | CSTs PathScan® Cleaved Caspase-3 (Asp175) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of cleaved caspase-3 protein. A total Caspase-3 Antibody has been coated onto the microwells. After incubation with cell lysates, the caspase-3 (cleaved and uncleaved) protein is captured by the coated antibody. Following extensive washing, A biotinylated Cleaved Caspase-3 Antibody is added to detect the captured cleaved caspase-3 protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of cleaved caspase-3 protein. Antibodies in kit are custom formulations specific to kit. CST的PathScan® Cleaved Caspase-3 (Asp175) Sandwich ELISA Kit采用了固相夹心酶联免疫吸附方法（ELISA）检测内源性水平切割的caspase-3蛋白。微孔中包被了total Caspase-3 Antibody。细胞裂解液孵育后，caspase-3蛋白（切割和未切割的）被包被的抗体捕获。充分洗涤后，添加biotinylated Cleaved Caspase-3 Antibody检测捕获的切割的caspase-3蛋白。再用HRP-linked streptavidin识别结合的抗体。添加HRP底物TMB用于显色。测得的吸光值与切割的caspase-3蛋白的量成正比。试剂盒中的抗体为该试剂盒所特制。 |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Cleaved Caspase-3 (Asp175) Sandwich ELISA Kit detects endogenous levels of cleaved caspase-3 protein in human cell lines. Using this ELISA kit #7190, a significant induction of cleaved caspase-3 at aspartic acid 175 can be seen in staurosporine-treated HeLa cells, while the level of total caspase-3, analyzed by western blot using Caspase-3 Antibody #9662, remains unchanged (Figure 1). Jurkat cells treated with etoposide show similar results (data not shown). 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 | Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires aspartic acid at the P1 position (2). Caspase-3 (CPP-32，Apoptain，Yama，SCA-1)是一个凋亡的关键效应执行者，因为它对许多关键的蛋白起着部分或全部的裂解作用，比如多聚核酶(ADP-ribose)的聚合酶(PARP)（1）。Caspase-3的活化需要经过其未激活的的酶原形式变为活化的p17 和 p12片段的蛋白水解过程。Caspase-3的裂解需要p1位存在天门冬氨酸（2）。 |
| 存放说明 | 4C |
| 参考文献 | Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4. Nicholson, D.W. et al. (1995) Nature 376, 37-43. Steiner, P. et al. (2007) Clin Cancer Res 13, 1540-51. |
Figure 2: Linear relationship between protein concentration of staurosporine-treated HeLa cell lysates and kit assay optical density readings. HeLa cells (90% confluence) were treated with staurosporine (1 uM), and lysed after incubation at 37ºC for 3 hours. 图2：星形孢菌素处理的Hela细胞裂解液蛋白浓度和试剂盒测得光密度读数之间的线性关系。星形孢菌素（1 uM）处理HeLa细胞（90％铺满），37℃下进行3小时的孵育后裂解。 | |
Figure 1: Treatment of HeLa cells with staurosporine stimulates cleavage of caspase-3 protein, detected by PathScan® Cleaved Caspase-3 (Asp175) Sandwich ELISA Kit #7190 (top figure), but does not affect the level of total caspase-3 protein detected by western blot (bottom figure), using Cleaved Caspase-3 (Asp175) Antibody #9661 (upper panel) or Caspase-3 Antibody #9662 (lower panel). 图1：PathScan® Cleaved Caspase-3 (Asp175) Sandwich ELISA Kit #7190检测到星形孢菌素处理的HeLa细胞中裂解caspase-3蛋白的含量（顶图），但不影响免疫印迹检测到的总caspase-3蛋白的水平（底图），使用抗体为Cleaved Caspase-3 (Asp175) Antibody #9661 （上图）或Caspase-3 Antibody #9662 （下图）。 |
