Abstract
In addition to methods aimed at the study of mitochondrial function in-situ, a full understanding of mitochondrial function requires their purification from cells or tissues under specific physiological or pathological conditions. This protocol illustrates a sequential procedure to obtain functional mitochondria with high yield from mice brain tissue. Mitochondria obtained with this method can be used to assess different mitochondrial parameters, including oxygen consumption, membrane potential and calcium retention capacity.
Keywords: Mitochondria, Brain, Respiration, Bioenergetics, Mice
Materials and Reagents
Equipment
Procedure
Notes:
Recipes
Acknowledgments
We thank Dr. Jorgina Satrustegui, in whose laboratory our previous work was carried out, and Dr. Araceli del Arco, for constant help, guidance, support and critical comments.
References
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Dear Hua Zhu,In our experiments (mainly respiratory assays or calcium retention capacity assays), we added 100 micromolar digitonin to the assay buffer. Digitonin was present during the whole experiment.Hope that helps. Best,Javier