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Hybrid Clear/Blue Native Electrophoresis for the Separation and Analysis of Mitochondrial Respiratory Chain Supercomplexes
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Hybrid Clear/Blue Native Electrophoresis for the Separation and Analysis of Mitochondrial Respiratory Chain Supercomplexes

Hybrid Clear/Blue Native Electrophoresis for the Separation and Analysis of Mitochondrial Respiratory Chain Supercomplexes

DOI:
10.3791/59294-v

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11:25 min

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May 19, 2019

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1Interdisciplinary School of Health Science, Faculty of Health Sciences and Department of Cellular and Molecular Medicine, Faculty of Medicine,University of Ottawa

Chapters

  • 00:04Title
  • 01:38Supercomplex Extraction
  • 02:36Gradient Gel Casting and Electrophoresis
  • 06:50In-gel Activity for Complexes I, II, IV and V
  • 08:24Results
  • 10:01Conclusion

Summary

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Here we present a protocol to extract, resolve and identify mitochondrial supercomplexes which minimizes exposure to detergents and Coomassie Blue. This protocol offers an optimal balance between resolution, and preservation of enzyme activities, while minimizing the risk of losing labile protein-protein interactions.

Tags

Mitochondrial Respiratory Chain SupercomplexesBlue Native PAGEClear Native PAGEHybrid Clear/blue Native ElectrophoresisMitochondrial IsolationDigitonin ExtractionGradient Gel Electrophoresis

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