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  • Measurement of mitochondrial respiratory chain enzymatic activities in Drosophila melanogaster samples.

Measurement of mitochondrial respiratory chain enzymatic activities in Drosophila melanogaster samples.

STAR protocols (2022-04-29)
Michele Brischigliaro, Elena Frigo, Erika Fernandez-Vizarra, Paolo Bernardi, Carlo Viscomi
摘要

Mitochondrial respiratory chain (MRC) dysfunction is linked to mitochondrial disease as well as other common conditions such as diabetes, neurodegeneration, cancer, and aging. Thus, the evaluation of MRC enzymatic activities is fundamental for diagnostics and research purposes on experimental models. Here, we provide a verified and reliable protocol for mitochondria isolation from various D. melanogaster samples and subsequent measurement of the activity of MRC complexes I-V plus citrate synthase (CS) through UV-VIS spectrophotometry. For complete details on the use and execution of this protocol, please refer to Brischigliaro et al. (2021).

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Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
鱼藤酮, ≥95%
Sigma-Aldrich
细胞色素 C 来源于马心脏, ≥95% based on Mol. Wt. 12,384 basis
Sigma-Aldrich
L-乳酸脱氢酶 来源于兔肌肉, Type II, ammonium sulfate suspension, 800-1,200 units/mg protein
Sigma-Aldrich
丙酮酸激酶 来源于兔肌肉, Type II, ammonium sulfate suspension, 350-600 units/mg protein
Sigma-Aldrich
乙二胺四乙酸 二钠盐 二水合物, BioUltra, 98.5-101.5%