Advanced

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 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)
Please use this url to cite or link to this publication:
author
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
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
2011-07-01 09:05:40
date last changed
2017-07-30 03:23:23
@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},
  volume       = {34},
  year         = {2011},
}