|
|||||
|
|
Blue native electrophoresis to study mitochondrial complex I in C. elegans |
|
| Authors: | Daniela van den Ecker Olaf Bossinger Leo G Nijtmans |
| Institution: | a Department of General Pediatrics, University Children’s Hospital, Heinrich-Heine-University, Düsseldorf, Germany b Nijmegen Centre for Mitochondrial Disorders, Department of Pediatrics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands c Institute ofMolecular andCellularAnatomy, RWTH Aachen University, Aachen, Germany |
| Abstract: | Blue native polyacrylamide gel electrophoresis (BN-PAGE) is an essential tool for investigating mitochondrial respiratory chain complexes. However, with current BN-PAGE protocols for Caenorhabditis elegans (C. elegans), large worm amounts and high quantities of mitochondrial protein are required to yield clear results. Here, we present an efficient approach to isolate mitochondrial complex I (NADH:ubiquinone oxidoreductase) from C. elegans, grown on agar plates. We demonstrate that considerably lower amounts of mitochondrial protein are sufficient to isolate complex I and to display clear in-gel activity results. Moreover, we present the first complex I assembly profile for C. elegans, obtained by two-dimensional BN/SDS-PAGE. |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ | |