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Erschienen in:

01.06.2011 | Original Article

Complex III staining in blue native polyacrylamide gels

verfasst von: Joél Smet, Boel De Paepe, Sara Seneca, Willy Lissens, Heike Kotarsky, Linda De Meirleir, Vineta Fellman, Rudy Van Coster

Erschienen in: Journal of Inherited Metabolic Disease | Ausgabe 3/2011

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Abstract

For more than a decade now blue native polyacrylamide gel electrophoresis (BN-PAGE) has been used for the study of the oxidative phosphorylation (OXPHOS) complexes. Catalytic activities of complexes I, II, IV and V can be assessed, after separation by gel electroforesis, by incubation of the BN-PAGE gel in specific staining solutions. However, until now, a reliable staining method for testing ubiquinol cytochrome c oxidoreductase (complex III) activity by BN-PAGE gel techniques was not available. In addition, spectrophotometric methods currently in use for detection of complex III deficiency in patients are not very sensitive. Here, we describe a newly developed diagnostic method for visualization of complex III activity by direct in-gel evaluation of ubiquinol cytochrome oxidoreductase activity. We validated the method by reporting the results in six patients with previously characterised complex III defects.

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Titel: Complex III staining in blue native polyacrylamide gels
verfasst von: Joél Smet
Boel De Paepe
Sara Seneca
Willy Lissens
Heike Kotarsky
Linda De Meirleir
Vineta Fellman
Rudy Van Coster
Publikationsdatum: 01.06.2011
Verlag: Springer Netherlands
Erschienen in: Journal of Inherited Metabolic Disease / Ausgabe 3/2011
Print ISSN: 0141-8955
Elektronische ISSN: 1573-2665
DOI: https://doi.org/10.1007/s10545-011-9315-7

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