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Quantification of Protein Complexes by Blue Native Electrophoresis

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Membrane Biogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1033))

Abstract

Blue native electrophoresis (BNE) is a long established method for the analysis of native protein complexes. Applications of BNE range from investigating subunit composition, stoichiometry, and assembly of single protein complexes to profiling of whole complexomes. BNE is an indispensible tool to diagnostically analyze cells and tissues from patients with mitochondrial disorders or model organisms. Since functional proteomic studies often require quantification of protein complexes, we describe here different quantification methods subsequent to protein complex separation by BNE.

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Acknowledgments

We are grateful for excellent technical assistance from Ilka Siebels. We thank Thilo Borchardt, Benno Jungblut, and Thomas Braun from Max Planck Institute for Heart and Lung Research (Bad Nauheim) for kindly providing newt and zebrafish samples. Many thanks to Arcangela Iuso from the Institute of Human Genetics (Helmholtz Center Munich) for wildtype flies (D.m.) and Stefan Dröse from the Molecular Bioenergetics group (Frankfurt) for mitochondria from Y.l. We thank Hermann Schägger for many helpful discussions and antibodies. The work was supported by the Bundesministerium für Bildung und Forschung (BMBF 01GM1113B; mitoNET—Deutsches Netzwerk für mitochondriale Erkrankungen) and by the Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 815 (Projects Z1 (Redox-Proteomics), A02 and A10), and Excellence Initiative (EXC 115).

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Authors and Affiliations

  1. Department of Molecular Hematology, University of Frankfurt Medical School, Frankfurt, Germany

    Juliana Heidler

  2. Molecular Bioenergetics, Medical School, Goethe-University, Frankfurt, Germany

    Valentina Strecker, Lea Bleier & Ilka Wittig

  3. Institute of Biochemistry, Goethe-University, Frankfurt, Germany

    Florian Csintalan

Authors
  1. Juliana Heidler
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  2. Valentina Strecker
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  3. Florian Csintalan
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  4. Lea Bleier
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  5. Ilka Wittig
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Editor information

Editors and Affiliations

  1. Interfaculty Institute of Biochemistry, University of Tuebingen, Tuebingen, Germany

    Doron Rapaport

  2. Cell Biology, University of Kaiserslautern, Kaiserslautern, Germany

    Johannes M. Herrmann

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Heidler, J., Strecker, V., Csintalan, F., Bleier, L., Wittig, I. (2013). Quantification of Protein Complexes by Blue Native Electrophoresis. In: Rapaport, D., Herrmann, J. (eds) Membrane Biogenesis. Methods in Molecular Biology, vol 1033. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-487-6_23

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  • DOI: https://doi.org/10.1007/978-1-62703-487-6_23

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-486-9

  • Online ISBN: 978-1-62703-487-6

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