Caspase protocols in mice.
Caspase protocols in mice.
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小鼠中的caspase方案。
DOI：
10.1007/978-1-4939-0357-3_9
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Kaushal, Gur P
中科院分区：
文献类型：
--
作者：
Kaushal, Varsha;Herzog, Christian;Haun, Randy S;Kaushal, Gur P
半胱天冬酶蛋白酶家族的成员是进化上保守的半胱氨酸蛋白酶，其作为细胞凋亡途径的中央执行者发挥关键作用。自半胱天冬酶被发现以来，人们开发了许多检测其活化的方法，并广泛应用于基础和临床研究。在小鼠组织中，可以通过切割胱天蛋白酶特异性合成底物和通过组织提取物的蛋白质印迹分析检测切割的胱天蛋白酶来监测胱天蛋白酶活化。在组织切片中，活性半胱天冬酶可以通过使用活性半胱天冬酶的特异性抗体进行免疫染色来检测。此外，在迄今为止已知的无数种半胱天冬酶特异性底物中，可以通过免疫染色以及组织提取物的蛋白质印迹在组织切片中监测由半胱天冬酶从底物如PARP、核纤层蛋白A和细胞角蛋白-18产生的切割片段。通常，应使用一种以上的方法来确定小鼠组织中半胱天冬酶激活的检测。
Members of the caspase family of proteases are evolutionarily conserved cysteine proteases that play a crucial role as the central executioners of the apoptotic pathway. Since the discovery of caspases, many methods have been developed to detect their activation and are widely used in basic and clinical studies. In a mouse tissue, caspase activation can be monitored by cleavage of caspase-specific synthetic substrates and by detecting cleaved caspase by western blot analysis of the tissue extract. In tissue sections, active caspase can be detected by immunostaining using specific antibodies to the active caspase. In addition, among the myriads of caspase-specific substrates known so far, cleaved fragments produced by caspases from the substrates such as PARP, lamin A, and cytokeratin-18 can be monitored in tissue sections by immunostaining as well as western blots of tissue extracts. In general, more than one method should be used to ascertain detection of activation of caspases in a mouse tissue.