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Complex III staining in blue native polyacrylamide gels

Smet, Joel ; De Paepe, Boel ; Seneca, Sara ; Lissens, Willy ; Kotarsky, Heike LU ; De Meirleir, Linda ; Fellman, Vineta LU orcid and Van Coster, Rudy (2011) In Journal of Inherited Metabolic Disease 34(3). p.741-747
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... (More)
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. (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Inherited Metabolic Disease
volume
34
issue
3
pages
741 - 747
publisher
Springer
external identifiers
  • wos:000291360100022
  • scopus:79959780054
  • pmid:21484424
ISSN
0141-8955
DOI
10.1007/s10545-011-9315-7
language
English
LU publication?
yes
id
871c5aeb-ae67-4cc7-94f4-9927b3022cd2 (old id 1985345)
date added to LUP
2016-04-01 10:52:58
date last changed
2022-02-10 06:55:52
@article{871c5aeb-ae67-4cc7-94f4-9927b3022cd2,
  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, Joel 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}},
  language     = {{eng}},
  number       = {{3}},
  pages        = {{741--747}},
  publisher    = {{Springer}},
  series       = {{Journal of Inherited Metabolic Disease}},
  title        = {{Complex III staining in blue native polyacrylamide gels}},
  url          = {{http://dx.doi.org/10.1007/s10545-011-9315-7}},
  doi          = {{10.1007/s10545-011-9315-7}},
  volume       = {{34}},
  year         = {{2011}},
}