Apoptosis-associated caspase activation assays
Apoptosis-associated caspase activation assays
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DOI：
10.1016/j.ymeth.2007.11.005
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发表时间：
2008-03-01
期刊：
影响因子：
4.8
通讯作者：
Earnshaw, William C.
中科院分区：
文献类型：
--
作者：
Kaufmann, Scott H.;Lee, Sun-Hee;Earnshaw, William C.
半胱氨酸氨基转移酶是一种天冬氨酸导向的半胱氨酸水解酶，它能裂解一组不同的细胞内底物，从而促进细胞凋亡的各种表现。这些蛋白水解酶被合成为不活跃的前体，并在广泛的生理和病理刺激诱导的信号转导下被激活。半胱氨酸天冬氨酸酶的激活可以通过测定催化活性、免疫印迹切割底物、使用构象敏感抗体进行免疫标记、亲和标记以及随后的流式细胞术或配基印迹来检测。在这里，我们描述了每一种分析方法，确定了这些分析方法的最新改进，并概述了每种方法的优点和局限性。(C)2007 Elsevier Inc.保留所有权利。
Caspases are aspartate-directed cysteine proteases that cleave a diverse group of intracellular substrates to contribute to various manifestations of apoptosis. These proteases are synthesized as inactive precursors and are activated as a consequence of signaling induced by a wide range of physiological and pathological stimuli. Caspase activation can be detected by measurement of catalytic activity, immunoblotting for cleavage of their substrates, immunolabeling using conformation-sensitive antibodies or affinity labeling followed by flow cytometry or ligand blotting. Here we describe methods for each of these assays, identify recent improvements in these assays and outline the strengths and limitations of each approach. (C) 2007 Elsevier Inc. All rights reserved.