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Simultaneous measurement of mitochondrial respiration and ATP production in tissue homogenates and calculation of effective P/O ratios ArchiMer
Salin, Karine; Villasevil, Eugenia M.; Auer, Sonya K.; Anderson, Graeme J.; Selman, Colin; Metcalfe, Neil B.; Chinopoulos, Christos.
The use of tissue homogenate has greatly aided the study of the functioning of mitochondria. However, the amount of ATP produced per oxygen molecule consumed, that is, the effective P/O ratio, has never been measured directly in tissue homogenate. Here we combine and refine existing methods previously used in permeabilized cells and isolated mitochondria to simultaneously measure mitochondrial ATP production (JATP) and oxygen consumption (JO(2)) in tissue homogenate. A major improvement over existing methods is in the control of ATPases that otherwise interfere with the ATP assay: our modified technique facilitates simultaneous measurement of the rates of uncorrected ATP synthesis and of ATP hydrolysis, thus minimizing the amount of tissue and time needed....
Tipo: Text Palavras-chave: ATPase; Fluorescence; Magnesium green; Oxidative phosphorylation; Oxygraph.
Ano: 2016 URL: https://archimer.ifremer.fr/doc/00414/52500/53300.pdf
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