Detection of Uncoupling Protein-2 (UCP2) As a Mitochondrial Modulator of Apoptosis
Detection of Uncoupling Protein-2 (UCP2) As a Mitochondrial Modulator of Apoptosis
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DOI：
10.1007/978-1-60327-017-5_15
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发表时间：
2009-01-01
期刊：
影响因子：
--
通讯作者：
Baffy, Gyorgy
中科院分区：
文献类型：
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作者：
Derdak, Zoltan;Garcia, Tamako A.;Baffy, Gyorgy
解偶联蛋白2（Uncoupling protein-2，UCP 2）是近年来发现的一种线粒体活性氧（reactive oxygen species，ROS）的分子传感器和抑制剂，在不同的细胞系统中对细胞凋亡起着重要的调控作用。许多研究小组遇到的一个很大的技术困难是可靠地检测内源或外源表达的UCP 2蛋白。本章的目的是向读者介绍我们多年来成功用于检测各种小鼠和人类标本中UCP 2蛋白的技术。这些技术包括线粒体分离和亚线粒体分级分离，然后进行Western印迹和UCP 2免疫组织化学。我们发现，样品制备是成功的关键，它允许人们使用市售抗体产生相关和重要的数据。
There is an increasing evidence that Uncoupling protein-2 (UCP2), a recently identified molecular sensor and Suppressor of mitochondrial reactive oxygen species (ROS), plays an important role in regulating apoptosis in different cell systems. A great technical difficulty that many groups have encountered is the reliable detection of endogenously or exogenously expressed UCP2 protein. The goal of this chapter is to introduce the reader to techniques that we have successfully used over the years to detect UCP2 protein in various Mouse and human specimens. These techniques include mitochondrial isolation and submitochondrial fractionation followed by Western blotting and UCP2 immunohistochemistry. We find that sample preparation is a key to success and it allows one to produce relevant and important data using commercially available antibodies.