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Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting
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Biologia
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JoVE Journal Biologia
Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting

Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting

DOI:
10.3791/59269-v

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07:55 min

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February 12, 2019

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1Research Programs Unit, Molecular Neurology,University of Helsinki

Capitoli

  • 00:04Titolo
  • 00:25Preparation of Mitochondrial Lysates
  • 02:40Preparation of Gradient Gel for BN-PAGE
  • 05:32Blue Native Gel Electrophoresis
  • 06:40Results: Investigating the Assembly of Mitochondrial OXPHOS Complexes Using BN-PAGE
  • 07:16Conclusion

Summary

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This protocol demonstrates the analysis of mitochondrial respiratory chain complexes by blue native polyacrylamide gel electrophoresis. Here the method applied to cultured human cells is described.

Tags

MitochondrialRespiratory ChainComplexesCulturedHumanCellsBlue NativePolyacrylamideGelElectrophoresisImmunoblottingOxidativePhosphorylationSystemIn-tactComplexesAssemblyPBSDigitoninProteaseInhibitorMitochondrialBufferLorealMountGradientGelPAGE

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