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Complex III staining in blue native polyacrylamide gels

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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.
Keywords
OXIDATIVE-PHOSPHORYLATION, LETHAL METABOLIC-DISORDER, GRACILE SYNDROME, IRON OVERLOAD, BCS1L GENE, DEFICIENCY, MUTATION, ELECTROPHORESIS, ASSAYS

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Chicago
Smet, Joél, Boel De Paepe, Sara Seneca, Willy Lissens, Heike Kotarsky, Linda De Meirleir, Vineta Fellman, and Rudy Van Coster. 2011. “Complex III Staining in Blue Native Polyacrylamide Gels.” Journal of Inherited Metabolic Disease 34 (3): 741–747.
APA
Smet, Joél, De Paepe, B., Seneca, S., Lissens, W., Kotarsky, H., De Meirleir, L., Fellman, V., et al. (2011). Complex III staining in blue native polyacrylamide gels. JOURNAL OF INHERITED METABOLIC DISEASE, 34(3), 741–747.
Vancouver
1.
Smet J, De Paepe B, Seneca S, Lissens W, Kotarsky H, De Meirleir L, et al. Complex III staining in blue native polyacrylamide gels. JOURNAL OF INHERITED METABOLIC DISEASE. 2011;34(3):741–7.
MLA
Smet, Joél, Boel De Paepe, Sara Seneca, et al. “Complex III Staining in Blue Native Polyacrylamide Gels.” JOURNAL OF INHERITED METABOLIC DISEASE 34.3 (2011): 741–747. Print.
@article{1233553,
  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.},
  author       = {Smet, Jo{\'e}l and De Paepe, Boel and Seneca, Sara and Lissens, Willy and Kotarsky, Heike and De Meirleir, Linda and Fellman, Vineta and Van Coster, Rudy},
  issn         = {0141-8955},
  journal      = {JOURNAL OF INHERITED METABOLIC DISEASE},
  keyword      = {OXIDATIVE-PHOSPHORYLATION,LETHAL METABOLIC-DISORDER,GRACILE SYNDROME,IRON OVERLOAD,BCS1L GENE,DEFICIENCY,MUTATION,ELECTROPHORESIS,ASSAYS},
  language     = {eng},
  number       = {3},
  pages        = {741--747},
  title        = {Complex III staining in blue native polyacrylamide gels},
  url          = {http://dx.doi.org/10.1007/s10545-011-9315-7},
  volume       = {34},
  year         = {2011},
}

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