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Free Flow Electrophoresis for Separation of Native Membrane Protein Complexes

  • Lutz Andreas Eichacker
  • Gerhard Weber
  • Ute Sukop-Köppel
  • Robert Wildgruber
Part of the Methods in Molecular Biology book series (MIMB, volume 1295)

Abstract

This chapter describes the technology of free flow electrophoresis (FFE) and protocols to separate membrane protein complexes for proteome analysis. FFE is a highly versatile technology applied in the field of protein analysis. It is superior to native PAGE due to its fast continuous processing of sample at high resolution. Additionally, the dynamic separation range from ions, peptides, to proteins, protein complexes, up to organelles, and whole cells makes it the method of choice in the analysis of proteins. FFE is carried out in an aqueous medium without inducing any solid matrix, such as acrylamide, so that it simplifies the analysis of protein complexes for the downstream analysis. Here, we describe the novel zone electrophoresis interval method (IZE-FFE) for separation of protein complexes from the thylakoid membrane of Arabidopsis thaliana by charge only. Protein complexes isolated by IZE FFE were characterized according to molecular weight by Blue Native PAGE and were proteins stained with coomassie.

Key words

Free flow electrophoresis Thylakoid membrane Protein complexes Blue Native PAGE 

Abbreviations

BN

Blue Native PAGE

FFE

Free flow electrophoresis

HPMC

Hydroxy-propyl-methylcellulose

iZE

Interval zone electrophoresis

PAGE

Polyacrylamide gel electrophoresis

ZE

Zone electrophoresis

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Lutz Andreas Eichacker
    • 1
  • Gerhard Weber
    • 2
  • Ute Sukop-Köppel
    • 2
  • Robert Wildgruber
    • 2
  1. 1.Center of Organelle Research (CORE)University of StavangerStavangerNorway
  2. 2.FFE Service GmbHFeldkirchenGermany

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