Caspase-3 Control Cell Extracts

• 产品编号：CST-9663S      品牌：CST       原厂货号：9663S
• 产品分类：蛋白质表达与纯化 > 蛋白质样本制备 > 细胞组织粗提物与裂解物
• 应用分类：

描述：

未处理的Jurkat 对照细胞提取物：未经处理的Jurkat细胞溶解在Chaps细胞提取物缓冲液中，产生一种胞质段，作为caspase裂解的阴性对照。用 Cytochrome c 处理的 Jurkat 细胞提取物：未处理的 Jurkat 细胞 溶解在 Chaps 细胞提取物缓冲液中，产生一个细胞质片段。然后，在体外用 Cytochrome c 处理后的提取物可以作为 caspase 裂解产物的阳性对照。Western Blots：建议电泳加样之前先对对照细胞提取物煮沸两分钟。每个泳道加入10μl的Jurkat对照细胞提取物。Caspase-3 (CPP-32, Apoptain, Yama, SCA-1)是一个凋亡的关键执行者，因为它对许多关键的蛋白起着部分或全部的裂解作用，比如核酶多聚(ADP-ribose)聚合酶(PARP)（1）。caspase-3的活化需要经过其未激活的的酶原转变成活化的 p17 和 p12 片段的蛋白水解过程。caspase-3的剪切需要天门冬氨酸位于p1位点（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.

原厂资料：

Description

Untreated Jurkat Control Cell Extracts: Untreated Jurkat cells are lysed in Chaps cell extract buffer and a cytoplasmic fraction is generated to serve as a negative control for caspase cleavage. 
Cytochrome c Treated Jurkat Cell Extracts: Untreated Jurkat cells are lysed in Chaps cell extract buffer and a cytoplasmic fraction is generated. Extracts are then treated with cytochrome c in vitro to generate a positive control for caspase cleavage.

Applications

Western Blots: CST recommends boiling control cell extracts for two minutes prior to loading. Load 10 µl of each Jurkat control cell extract per lane.

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.

注意事项：

Storage: Supplied in SDS Sample Buffer: 62.5 mM Tris-HCl
(pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol,50 mM DTT,
0.01% w/v bromophenol blue or phenolred. Store at –20°C, or at
–80°C for long-term storage.