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Updated: Oct 23, 2025
Isolation of Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High Throughput Microplate Respiratory Measurements
Published on: November 13, 2015
An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse
Cristina Sánchez-González1, Laura Formentini1,2,3
1Departamento de Biología Molecular, Centro de Biología Molecular "Severo Ochoa" (CBMSO), Universidad Autónoma de Madrid, c/ Nicolas Cabrera 1, 28049, Madrid, Spain.
Abstract:
Depending on metabolic requirements, skeletal muscle mitochondria integrate O2 consumption and ATP production with lipid, glucose, or amino acid metabolism. Free fatty acids (FFAs) are the main source of energy during rest and mild-intensity exercise. We present a detailed protocol for measuring FFA-β-oxidation coupled with O2 respiration by a Clark-type electrode in isolated mitochondria from mouse soleus oxidative muscle. We optimized the procedure, including buffer composition, protease treatment, and quantifiable parameters (P/O, Phosphate/Oxygen Ratio; OCR, Oxygen Consumption Rate; RCR,Respiration Control Rate; OSR, Oligomycin Sensitive Respiration). For complete details on the use and execution of this protocol, please refer to Sanchez-Gonzalez et al. (2020).
