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.
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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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Eichacker, L.A., Weber, G., Sukop-Köppel, U., Wildgruber, R. (2015). Free Flow Electrophoresis for Separation of Native Membrane Protein Complexes. In: Posch, A. (eds) Proteomic Profiling. Methods in Molecular Biology, vol 1295. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2550-6_29
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DOI: https://doi.org/10.1007/978-1-4939-2550-6_29
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2549-0
Online ISBN: 978-1-4939-2550-6
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