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- Publisher
- Springer Journals
- Copyright
- Copyright © 2012 by Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.
- Subject
- Life Sciences; Life Sciences, general; Biological Techniques; Analytical Chemistry; Microarrays; Computational Biology/Bioinformatics; Organic Chemistry
- ISSN
- 1754-2189
- eISSN
- 1750-2799
- DOI
- 10.1038/nprot.2012.058
- Publisher site
- See Article on Publisher Site
Abstract
The assessment of mitochondrial respiratory chain (RC) enzymatic activities is essential for investigating mitochondrial function in several situations, including mitochondrial disorders, diabetes, cancer, aging and neurodegeneration, as well as for many toxicological assays. Muscle is the most commonly analyzed tissue because of its high metabolic rates and accessibility, although other tissues and cultured cell lines can be used. We describe a step-by-step protocol for a simple and reliable assessment of the RC enzymatic function (complexes I–IV) for minute quantities of muscle, cultured cells and isolated mitochondria from a variety of species and tissues, by using a single-wavelength spectrophotometer. An efficient tissue disruption and the choice for each assay of specific buffers, substrates, adjuvants and detergents in a narrow concentration range allow maximal sensitivity, specificity and linearity of the kinetics. This protocol can be completed in 3 h.
Journal
Nature Protocols – Springer Journals
Published: May 31, 2012