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Blue-Native Electrophoresis to Study the OXPHOS Complexes

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Mitochondrial Gene Expression

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2192))

Abstract

Blue-native polyacrylamide gel electrophoresis (BN-PAGE) is a technique optimized for the analysis of the five components of the mitochondrial oxidative phosphorylation (OXPHOS) system. BN-PAGE is based on the preservation of the interactions between the individual subunits within the integral complexes. To achieve this, the complexes are extracted from the mitochondrial inner membrane using mild detergents and separated by electrophoresis in the absence of denaturing agents. The electrophoretic procedures can then be combined with a variety of downstream detection techniques. Since its development in the 1990s, BN-PAGE has been applied in the study of mitochondria from all kinds of organisms and extensive amounts of data have been produced using this technique, being key for the understanding of many aspects of OXPHOS physiopathology.

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Acknowledgements

We are grateful to Dr. José L. Capablo (Neurology Department, Miguel Servet University Hospital, Zaragoza, Spain) and Robert D.S. Pitceathly (MRC Centre for Neuromuscular Diseases, UCL Institute of Neurology and National Hospital for Neurology and Neurosurgery, London, UK) for providing pathological human samples. Our research was funded by Core Grant from the MRC (Grant MC_UU_00015/5), ERC Advanced Grant FP7-322424 and NRJ-Institut de France Grant to M.Z.; and by a “Miguel Servet” Grant (CP09/00156) from the Instituto de Salud Carlos III (Ministerio de Economía y Competitividad), “Marie Curie” European Reintegration Grant (PERG04-GA-2008-239372) and Association Française contre les Myopathies Trampoline Grant (AFM 14921) and Research Grant (AFM 16086) to E.F.-V.

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Authors and Affiliations

  1. MRC Mitochondrial Biology Unit, University of Cambridge, Cambridge, UK

    Erika Fernandez-Vizarra & Massimo Zeviani

Authors
  1. Erika Fernandez-Vizarra
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  2. Massimo Zeviani
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Correspondence to Erika Fernandez-Vizarra .

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Editors and Affiliations

  1. Medical Research Council Mitochondrial Biology Unit, University of Cambridge, Cambridge, UK

    Michal Minczuk

  2. Division of Molecular Metabolism, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Solna, Sweden, Institutet Laboratory, Max Planck Institute Biology of Ageing - Karolinska, Karolinska Institutet, Stockholm, Sweden

    Joanna Rorbach

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Fernandez-Vizarra, E., Zeviani, M. (2021). Blue-Native Electrophoresis to Study the OXPHOS Complexes. In: Minczuk, M., Rorbach, J. (eds) Mitochondrial Gene Expression. Methods in Molecular Biology, vol 2192. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0834-0_20

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  • DOI: https://doi.org/10.1007/978-1-0716-0834-0_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0833-3

  • Online ISBN: 978-1-0716-0834-0

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