An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus

STAR Protocols

Laura Formentini¹  Cristina Sánchez-González² 

[1] Corresponding author;Departamento de Biología Molecular, Centro de Biología Molecular “Severo Ochoa” (CBMSO), Universidad Autónoma de Madrid, c/ Nicolas Cabrera 1, 28049, Madrid, Spain;

关键词: cell biology;    cell isolation;    cell membrane;    cell separation/fractionation;    metabolism;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown