Mitochondrial ATP production measurement

After the measurement of ADP-stimulated respiration, the incubation buffer containing mitochondria was taken from the respiration chamber and immediately supplemented with the ATP Assay Mix (diluted 1 : 5) (Sigma, Aldrich). Mitochondrial ATP production after each respiration measurement was determined immediately and compared with ATP standards using a 96-well white plate and a spectrofluorometer (SpectraMax® M3, Molecular Devices, EUA) at 560 nm emission wavelength [27 (link)].

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Matta L., Fonseca T.S., Faria C.C., Lima-Junior N.C., De Oliveira D.F., Maciel L., Boa L.F., Pierucci A.P., Ferreira A.C., Nascimento J.H., Carvalho D.P, & Fortunato R.S. (2021). The Effect of Acute Aerobic Exercise on Redox Homeostasis and Mitochondrial Function of Rat White Adipose Tissue. Oxidative Medicine and Cellular Longevity, 2021, 4593496.

Publication 2021

ATP Assay Mix SpectraMax® M3

Assay Buffer Devices Mitochondrial Respiration

Corresponding Organization : Universidade Federal do Rio de Janeiro

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Variable analysis

independent variables

Measurement of ADP-stimulated respiration

dependent variables

Mitochondrial ATP production

control variables

Incubation buffer containing mitochondria
ATP Assay Mix (diluted 1 : 5)
Spectrofluorometer (SpectraMax® M3, Molecular Devices, EUA) at 560 nm emission wavelength

controls

ATP standards

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