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Anal Biochem. 2010 Dec 15;407(2):287-9. doi: 10.1016/j.ab.2010.08.009. Epub 2010 Aug 10.

Blue native electrophoresis to study mitochondrial complex I in C. elegans.

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1
Department of General Pediatrics, University Children's Hospital, Heinrich-Heine-University, Düsseldorf, Germany.

Abstract

Blue native polyacrylamide gel electrophoresis (BN-PAGE) is an essential tool for investigating mitochondrial respiratory chain complexes. However, with current BN-PAGE protocols for Caenorhabditis elegans (C. elegans), large worm amounts and high quantities of mitochondrial protein are required to yield clear results. Here, we present an efficient approach to isolate mitochondrial complex I (NADH:ubiquinone oxidoreductase) from C. elegans, grown on agar plates. We demonstrate that considerably lower amounts of mitochondrial protein are sufficient to isolate complex I and to display clear in-gel activity results. Moreover, we present the first complex I assembly profile for C. elegans, obtained by two-dimensional BN/SDS-PAGE.

PMID:
20705045
DOI:
10.1016/j.ab.2010.08.009
[Indexed for MEDLINE]
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