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. © 2014 Springer Science+Business Media, LLC.
CITATION STYLE
Arnold, J., Shapiguzov, A., Fucile, G., Rochaix, J. D., Goldschmidt-Clermont, M., & Eichacker, L. A. (2014). Separation of membrane protein complexes by native LDS-PAGE. Methods in Molecular Biology, 1072, 667–676. https://doi.org/10.1007/978-1-62703-631-3_46
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