PathScan? Cleaved Caspase-3 (Asp175) Sandwich ELISA Kit

• 产品编号：CST-7190C      品牌：CST       原厂货号：7190C
• 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 Caspase-3 (Asp175) Sandwich ELISA Kit能够检测内源性水平人细胞系切割的caspase-3蛋白。使用该ELISA试剂盒#7190能检测到星形孢菌素处理的HeLa细胞中Asp175位切割的caspase-3蛋白的显著减少，而通过western blot方法利用抗体Caspase-3 Antibody #9662测得的总的caspase-3水平不发生变化（如图1所示）。依托泊苷处理的Jurkat细胞表现出类似的结果（数据未示出）。本试剂盒可检测经内部测试确定的指定物种的蛋白质，同时也可以检测其他物种的同源蛋白。CST的PathScan® Cleaved 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。 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）。

1. Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4.
2. Nicholson, D.W. et al. (1995) Nature 376, 37-43.
3. Steiner, P. et al. (2007) Clin Cancer Res 13, 1540-51.

原厂资料：

Specificity / Sensitivity

CST's 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. 

Description

CST's 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.

Background

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). 

1. Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4.
2. Nicholson, D.W. et al. (1995) Nature 376, 37-43.
3. Steiner, P. et al. (2007) Clin Cancer Res 13, 1540-51.

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).