Complex III staining in blue native polyacrylamide gels
- Author
- Joél Smet (UGent) , Boel De Paepe (UGent) , Sara Seneca, Willy Lissens, Heike Kotarsky, Linda De Meirleir, Vineta Fellman and Rudy Van Coster (UGent)
- Organization
- 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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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-1233553
- MLA
- Smet, Joél, et al. “Complex III Staining in Blue Native Polyacrylamide Gels.” JOURNAL OF INHERITED METABOLIC DISEASE, vol. 34, no. 3, 2011, pp. 741–47, doi:10.1007/s10545-011-9315-7.
- APA
- Smet, J., De Paepe, B., Seneca, S., Lissens, W., Kotarsky, H., De Meirleir, L., … Van Coster, R. (2011). Complex III staining in blue native polyacrylamide gels. JOURNAL OF INHERITED METABOLIC DISEASE, 34(3), 741–747. https://doi.org/10.1007/s10545-011-9315-7
- Chicago author-date
- 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): 741–47. https://doi.org/10.1007/s10545-011-9315-7.
- Chicago author-date (all authors)
- 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. doi:10.1007/s10545-011-9315-7.
- 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.
- IEEE
- [1]J. Smet et al., “Complex III staining in blue native polyacrylamide gels,” JOURNAL OF INHERITED METABOLIC DISEASE, vol. 34, no. 3, pp. 741–747, 2011.
@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, Joél 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}}, keywords = {{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://doi.org/10.1007/s10545-011-9315-7}}, volume = {{34}}, year = {{2011}}, }
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