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Analyzing Supercomplexes of the Mitochondrial Electron Transport Chain with Native Electrophoresis, In-gel Assays, and Electroelution
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Biochemistry
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JoVE Journal Biochemistry
Analyzing Supercomplexes of the Mitochondrial Electron Transport Chain with Native Electrophoresis, In-gel Assays, and Electroelution

Analyzing Supercomplexes of the Mitochondrial Electron Transport Chain with Native Electrophoresis, In-gel Assays, and Electroelution

DOI:
10.3791/55738-v

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08:37 min

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June 01, 2017

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1Department of Pediatrics-Division Cardiology,University of Rochester

Chapters

  • 00:05Title
  • 00:58In-gel Assays for Cx-I and Cx-V
  • 03:19Electroelution
  • 05:59Results: Isolation, Visualization, amd Analysis of Mitochondrial Supercomplexes
  • 07:00Conclusion

Summary

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This protocol describes the separation of functional mitochondrial electron transport chain complexes (Cx) I-V and supercomplexes thereof using native electrophoresis to reveal information about their assembly and structure. The native gel can be subjected to immunoblotting, in-gel assays, and purification by electroelution to further characterize individual complexes.

Tags

Keywords: Mitochondrial Electron Transport ChainSupercomplexesNative ElectrophoresisIn-gel AssaysElectroelutionComplex-1 AssayComplex-5 AssayDetergentsNADHOligomycinNitro Blue TetrazoliumLead NitrateATPBioenergeticsPhysiological IntegrityMembrane Protein Complexes

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