Apoptosis-associated caspase activation assays

详细信息    查看全文

• 作者：Scott H. Kaufmann ; Sun-Hee Lee ; X. Wei Meng ; David A. Loegering ; Timothy J. Kottke ; Alex ; er J. Henzing ; S ; rine Ruchaud ; Kumiko Samejima ; William C. Earnshaw
• 关键词：Caspases ; Apoptosis ; Affinity labeling ; Enzymatic activity ; Fluorogenic substrate ; Flow cytometry ; Immunoblotting ; Immunofluorescence
• 刊名：Methods
• 出版年：2008
• 出版时间：March 2008
• 年：2008
• 卷：44
• 期：3
• 页码：262-272
• 全文大小：512 K

文摘

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.