Procaspase Antibody Sampler Kit

• 产品编号：CST-12742S      品牌：Cell Signaling Technology       原厂货号：12742S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

Procaspase Antibody Sampler 试剂盒中的每个抗体都能够检测其对应的内源性靶蛋白。Caspase-3 (8G10) Rabbit mAb 兔单抗能够检测caspase-3全长(35 kDa)或在Asp175裂解的caspase-3大片段(17/19 kDa)。Caspase-6 抗体能够检测全长caspase-6(35 kDa)和在Asp193 裂解的caspase-6的小亚基(15 kDa)。Caspase-7 (D2Q3L) Rabbit mAb 兔单抗能够检测全长(35 kDa)的Caspase-7以及在Asp198裂解产生的caspase-7小亚基(20kDa)。Caspase-8 (1C12) Mouse mAb 鼠单抗可以检测到caspase-8的全长(57 kDa),断裂产生的中部p43/p41以及p18片段。Caspase-9 (C9) Antibody能够检测到全长caspase-9，以及在Asp315和 Asp330裂解产生的片段。PARP Antibody能够检测全长PARP1(116 kDa)，以及被caspase在Asp214处裂解产生的大片段(89kDa)。Lamin A/C (4C11) Mouse mAb 鼠单抗能够检测全长lamin A和lamin C蛋白，也可以识别被caspase 水解产生的lamin A (50 kDa)和lamin C (41 kDa)的大片段。Caspase-9 (C9)抗体能够识别caspase-9的前体形式以及水解片段。其中的单克隆抗体由合成肽段免疫动物而制备产生的，这些肽段分别与人caspase-3蛋白(Asp175)附近氨基酸，人caspase-7蛋白Pro158附近氨基酸，人caspase-8的p18片段羧基端序列，重组人caspase-9或人lamin A蛋白一致。其中的多抗由合成的肽段免疫动物获得，肽段和PARP的capase-6识别位点或caspase切断位点附近的氨基酸残基序列一致，抗体由蛋白A和肽段亲和层析技术纯化得到。Procaspase Antibody Sampler试剂盒提供了一种经济的方式分析caspases的丰度和活性。试剂盒含有的一抗足以进行4次western blot实验的。

细胞凋亡是一个被调控的生理过程，导致细胞死亡。Caspases，是一个半胱氨酸蛋白酶家族，是细胞凋亡的中心调控因子。引发剂 caspases（包括8、9、10和12）被紧密耦合到促凋亡信号。一旦被激活，这些 caspases 切割并激活下游的效应 caspases（包括3、6和7），它们反过来切割细胞骨架和核蛋白质，如PARP、α-胞衬蛋白、DFF和 lamin A，并诱导细胞凋亡(1,2)。

Caspase-8 (FLICE、Mch5、MACH) 和Caspase-9 (ICE-LAP6、Mch6) 是起始caspases。CD95受体(Fas/APO-1)和肿瘤坏死因子受体1(TNFR1)激活caspase-8，导致caspase激活片段的释放，包括p18 和 p10(3-6)。细胞色素C从线粒体释放，并与procaspase-9 (47 kDa)/Apaf 1交联。此交联复合物促使caspase-9从无酶原活性转变成活化状态，并在Asp315断裂产生p53亚基。另一个断裂发生在Asp330，产生p37亚基，从而放大凋亡应激信号(7-11)。

Caspase-3 (CPP-32、Apoptain、Yama、SCA-1)、Caspase-6 (Mch2)和 Caspase-7 (CMH-1、Mch3、ICE-LAP3)是效应caspases(12-16)。Caspase-3的活化需要经过其未激活酶原形式变为活化的p17 和 p12片段的蛋白水解过程(17)。Procaspase-7被上游caspases在Asp23、Asp198和Asp206位点蛋白水解产生成熟的亚基(14,16)。Procaspase-6可以被caspase-3在Asp23、Asp179 和 Asp193位点水解产生活化的大(p18)和小(p11)亚基(7)。

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(18)。在体外该蛋白可被许多ICE样 caspases裂解(2,19)，在体内是caspase-3裂解的主要靶蛋白之一(17,20)。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(17,19)。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(12）。核纤层蛋白Lamin是核膜结构的组分，对维持正常的细胞功能非常重要，例如细胞周期控制、DNA修复和染色质组装（22-24）。

Lamin A/C能够被caspase-6切割，作为caspase-6激活的标志。在凋亡过程中，Lamin A/C能够被剪切成特异的大片段(41-50 kDa)和小片段 (28 kDa)（24,25）。Lamin的剪切导致细胞核的失调和细胞死亡（26,27）。

原厂资料：

Specificity / Sensitivity

Each antibody in the Procaspase Antibody Sampler Kit detects endogenous levels of its respective target. Caspase-3 (8G10) Rabbit mAb detects full-length (35 kDa) and the large fragment (17/19 kDa) of caspase-3 resulting from cleavage at Asp175. Caspase-6 Antibody detects both full length caspase-6 (35 kDa) and the small subunit (15 kDa) of caspase-6 resulting from cleavage at Asp193. Caspase-7 (D2Q3L) Rabbit mAb detects both the full-length (35 kDa) and the large subunit (20 kDa) of caspase-7 resulting from cleavage at Asp198. Caspase-8 (1C12) Mouse mAb detects full length (57 kDa), the cleaved intermediate p43/p41, and the p18 fragment of caspase-8. Caspase-9 (C9) Antibody detects full-length caspase-9, as well as the large fragments resulting from cleavage at Asp315 and Asp330. PARP Antibody detects full length PARP1 (116 kDa), as well as the large fragment (89 kDa) of PARP1 resulting from caspase cleavage at Asp214. Lamin A/C (4C11) Mouse mAb detects full-length lamin A and lamin C proteins, as well as the larger fragments of lamin A (50 kDa) and lamin C (41 kDa) resulting from caspase cleavage. Caspase-9 (C9) Antibody detects endogenous levels of the pro form of caspase-9 as well as cleaved fragments.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues adjacent to (Asp175) in human caspase-3 protein, residues surrounding Pro158 of human caspase-7 protein, the carboxy-terminal sequence of the p18 fragment of human caspase-8 protein, recombinant human caspase-9 protein or human lamin A protein.Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the cleavage site of caspase-6 or the caspase cleavage site in PARP. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Description

The Procaspase Antibody Sampler Kit provides an economical means to evaluate the abundance and activation of caspases. The kit contains enough primary antibody to perform at least four western blots per primary antibody.

Background

Apoptosis is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 2, 8, 9, 10 and 12) are closely coupled to proapoptotic signals, which include the FasL, TNF-α, and DNA damage. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6 and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF and lamin A, and induce apoptosis (1,2).Caspase-8 (FLICE, Mch5, MACH) and Caspase-9 (ICE-LAP6, Mch6) are initiator caspases. CD95 receptor (Fas/APO-1) and tumor necrosis factor receptor 1 (TNFR1) activate caspase-8, leading to the release of the caspase-8 active fragments, p18 and p10 (3-6). Cytochrome c released from the mitochondria associates with procaspase-9 (47 kDa)/Apaf 1. Apaf-1 mediated activation of caspase-9 involves intrinsic proteolytic processing resulting in cleavage at Asp315 and producing a p35 subunit. Another cleavage occurs at Asp330 producing a p37 subunit that can serve to amplify the apoptotic response (7-11).

Caspase-3 (CPP-32, Apoptain, Yama, SCA-1), Caspase-6 (Mch2), and Caspase-7 (CMH-1, Mch3, ICE-LAP3) are effector caspases (12-16). Activation of caspase-3 requires proteolytic processing of its inactive zymogen/proform into activated p17 and p12 subunits (17). Procaspase-7 is activated through proteolytic processing by upstream caspases at Asp23, Asp198, and Asp206 to produce the mature subunits (14,16). Procaspase-6 is cleaved by caspase-3 at Asp23, Asp179 and Asp193 to form active large (p18) and small (p11) subunits (7).

PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (18). This protein can be cleaved by many ICE-like caspases in vitro (2,19) and is one of the main cleavage targets of caspase-3 in vivo (17,20). 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) (17,19). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (21).

Lamins are nuclear membrane structural components that are important in maintaining normal cell functions such as cell cycle control, DNA replication, and chromatin organization (22-24). Lamin A/C is cleaved by caspase-6 and serves as a marker for caspase-6 activation. During apoptosis, lamin A/C is specifically cleaved into large (41-50 kDa) and small (28 kDa) fragments (24,25). The cleavage of lamins results in nuclear disregulation and cell death (26,27).

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibodies.