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Scientific article
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Separation of membrane protein complexes by native LDS-PAGE

Published inMethods in molecular biology, vol. 1072, p. 667-676
Publication date2014
Abstract

Gel electrophoresis has become one of the most important methods for the analysis of proteins and protein complexes in a molecular weight range of 1-10(7) kDa. The separation of membrane protein complexes remained challenging to standardize until the demonstration of Blue Native PAGE in 1991 [1] and Clear Native PAGE in 1994 [2]. We present a robust protocol for high-resolution separation of photosynthetic complexes from Arabidopsis thaliana using lithium dodecyl sulfate as anion in a modified Blue Native PAGE (LDS-PAGE). Here, non-covalently bound chlorophyll is used as a sensitive probe to characterize the assembly/biogenesis of the pigment-protein complexes essential for photosynthesis. The high fluorescence yield recorded from chlorophyll-binding protein complexes can also be used to establish the separation of native protein complexes as an electrophoretic standard.

Keywords
  • Native LDS-PAGE
  • Thylakoid membrane
  • Chlorophyll-binding protein complexes
  • Fluorescence
Citation (ISO format)
ARNOLD, Janine et al. Separation of membrane protein complexes by native LDS-PAGE. In: Methods in molecular biology, 2014, vol. 1072, p. 667–676. doi: 10.1007/978-1-62703-631-3_46
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ISSN of the journal1064-3745
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