Simplified method for concentration of mitochondrial membrane protein complexes.
Extracting and concentrating mitochondrial protein complexes from gel strips after blue native PAGE (BN-PAGE) can be daunting tasks using the traditional methods, such as electroelution, passive diffusion and centrifugal concentration. We present a simplified gel electrophoresis method to concentrate mitochondrial protein complexes with excellent recovery rate. Mitochondrial complex I present in a long gel strip from BN-PAGE can be easily concentrated into a 0.8 cm gel strip when a second BN-PAGE is performed with a Y-shaped gel and the addition of 0.01% n-dodecyl beta-D-maltoside and 0.001% SDS in the cathode buffer. Once completed, the concentrated protein complex in the gel strip is ready for SDS-PAGE or proteomic studies.
Yeh
Steve T
ST
Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, USA.
Angelos
Mark G
MG
Chen
Yeong-Renn
YR
D000818:Animals; D042967:Electron Transport Complex I / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005960:Glucosides / Q000737:chemistry; D051336:Mitochondrial Membranes / Q000737:chemistry*; D024101:Mitochondrial Proteins / Q000737:chemistry*; D051381:Rats; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
D005960:Glucosides; D024101:Mitochondrial Proteins; D012967:Sodium Dodecyl Sulfate; C040358:dodecyl maltoside; D042967:Electron Transport Complex I