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Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue
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Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue

Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue

DOI:
10.3791/52076-v

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09:27 min

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March 23, 2015

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, , , ,
1Chemistry Department,Elon University, 2Department of Neurobiology and Anatomy,Wake Forest School of Medicine, 3Neuroscience Graduate Program,Wake Forest School of Medicine, 4ALS Center Translational Science Unit,Wake Forest School of Medicine

Chapters

  • 00:05Title
  • 01:49Cell Culture
  • 02:24Isolation of Mitochondria from Cultured Cells
  • 04:58Mitochondria Isolation from Mouse Tissue
  • 06:26Assessment of Mitochondrial (Dys)function
  • 07:42Results: Cytokine Treatment Leads to Increased Cell Death and Decreased Membrane Potential in Mitochondria
  • 08:51Conclusion

Summary

Automatic Translation

Automatic Translation

Comparison of mitochondrial membrane potential between samples yields valuable information about cellular status. Detailed steps for isolating mitochondria and assessing response to inhibitors and uncouplers using fluorescence are described. The method and utility of this protocol are illustrated by use of a cell culture and animal model of cellular stress.

Tags

Keywords: Mitochondria IsolationMitochondrial FunctionCell CultureTMREMitochondrial UncouplersMitochondrial InhibitorsFluorescent Plate ReaderHigh-throughput AnalysisOrganelle-specific FunctionCell MetabolismEnergy ProductionApoptosis

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