Original Article

Journal of Inherited Metabolic Disease

, Volume 34, Issue 3, pp 741-747

Complex III staining in blue native polyacrylamide gels

  • Joél SmetAffiliated withDepartment of Pediatrics, Division of Pediatric Neurology and Metabolism, Ghent University Hospital
  • , Boel De PaepeAffiliated withDepartment of Pediatrics, Division of Pediatric Neurology and Metabolism, Ghent University Hospital
  • , Sara SenecaAffiliated withCenter for Medical Genetics, UZ Brussel and Reproduction and Genetics (REGE), Vrije Universiteit Brussel
  • , Willy LissensAffiliated withCenter for Medical Genetics, UZ Brussel and Reproduction and Genetics (REGE), Vrije Universiteit Brussel
  • , Heike KotarskyAffiliated withDepartment of Pediatrics, Clinical Sciences, Lund University
  • , Linda De MeirleirAffiliated withCenter for Medical Genetics, UZ Brussel and Reproduction and Genetics (REGE), Vrije Universiteit Brussel
  • , Vineta FellmanAffiliated withDepartment of Pediatrics, Clinical Sciences, Lund UniversityDepartment of Pediatrics, Clinical Sciences, University of Helsinki
  • , Rudy Van CosterAffiliated withDepartment of Pediatrics, Division of Pediatric Neurology and Metabolism, Ghent University Hospital Email author 

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

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.