Separation of membrane protein complexes by native LDS-PAGE.
Methods Mol Biol
; 1072: 667-76, 2014.
Article
em En
| MEDLINE
| ID: mdl-24136555
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Dodecilsulfato de Sódio
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Arabidopsis
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Proteínas de Arabidopsis
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Eletroforese em Gel de Poliacrilamida
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Proteínas de Membrana
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article