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Electrophoresis. 2010 Jun;31(12):1934-6. doi: 10.1002/elps.201000019.

Simplified method for concentration of mitochondrial membrane protein complexes.

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  • 1Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, USA.

Abstract

Extracting and concentrating mitochondrial protein complexes from gel strips after blue native PAGE (BN-PAGE) can be daunting tasks using the traditional methods, such as electroelution, passive diffusion and centrifugal concentration. We present a simplified gel electrophoresis method to concentrate mitochondrial protein complexes with excellent recovery rate. Mitochondrial complex I present in a long gel strip from BN-PAGE can be easily concentrated into a 0.8 cm gel strip when a second BN-PAGE is performed with a Y-shaped gel and the addition of 0.01% n-dodecyl beta-D-maltoside and 0.001% SDS in the cathode buffer. Once completed, the concentrated protein complex in the gel strip is ready for SDS-PAGE or proteomic studies.

PMID:
20503203
[PubMed - indexed for MEDLINE]
PMCID:
PMC3698224
Free PMC Article

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