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Over several years we have provided evidence that uncoupling protein 1 (UCP1) is present in thymus mitochondria. We have demonstrated the conclusive evidence for the presence of UCP1 in thymus mitochondria and we have been able to demonstrate a GDP-sensitive UCP1-dependent proton leak in non-phosphorylating thymus mitochondria. In this chapter, we show how to detect UCP1 in mitochondria isolated from whole thymus using immunoblotting. We show how to measure GDP-sensitive UCP1-dependent oxygen consumption in non-phosphorylating thymus mitochondria and we show that increased reactive oxygen species production occurs on addition of GDP to non-phosphorylating thymus mitochondria. We conclude that reactive oxygen species production rate can be used as a surrogate for detecting UCP1 catalyzed proton leak activity in thymus mitochondria.

Original publication




Journal article


Methods Mol Biol

Publication Date





123 - 135


Animals, Biological Transport, Cell Fractionation, Electrophoresis, Polyacrylamide Gel, Female, Guanosine Diphosphate, Hydrogen Peroxide, Immunoblotting, Ion Channels, Male, Mitochondria, Mitochondrial Proteins, Oxygen Consumption, Protons, Rats, Rats, Wistar, Thymus Gland, Uncoupling Protein 1