Summary
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.
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Acknowledgments
Experimental studies providing the basis of this work were supported by National Institutes of Health grants DK-61890 and RR-17695 to G.B. We thank Rose M. Tavares at the Department of Pathology, Rhode Island Hospital, for her help with immunohistochemistry studies. The authors of this article declare no financial conflict of interest.
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Derdak, Z., Garcia, T.A., Baffy, G. (2009). Detection of Uncoupling Protein-2 (UCP2) As a Mitochondrial Modulator of Apoptosis. In: Erhardt, P., Toth, A. (eds) Apoptosis. Methods in Molecular Biology, vol 559. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-017-5_15
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DOI: https://doi.org/10.1007/978-1-60327-017-5_15
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