A critical comparison between two classical and a kit-based method for mitochondria isolation.
Numerous protocols for isolation of mitochondria are available. Here, three methods for the isolation of intact mitochondria from mouse liver tissues are compared with regard to yield, purity and activity. Mitochondria were isolated by sucrose density gradient ultracentrifugation, free-flow electrophoresis or a commercially available kit-based method. Our analyses show that the sophisticated (and most expensive) free-flow electrophoresis method enables isolation of intact mitochondria with an enrichment of approximately 70%. Using the classical density centrifugation method is very laborious and time-consuming, but delivers about 57% intact mitochondria. Using standard laboratory equipment in a quick and simple procedure, the kit provides approximately 50% intact mitochondria, suitable for most standard investigations.
Institute of Clinical Biochemistry and Pathobiochemistry, German Diabetes Center at the Heinrich-Heine-University Düsseldorf, Leibniz Center for Diabetes Research, Düsseldorf, Germany.
Sonja
S
Hartwig
Christian
C
Feckler
Stefan
S
Lehr
Katrin
K
Wallbrecht
Heike
H
Wolgast
Dirk
D
Müller-Wieland
Jörg
J
Kotzka
D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation / Q000379:methods*; D004586:Electrophoresis / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D051379:Mice; D008930:Mitochondria, Liver; D040901:Proteomics; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013385:Succinate Dehydrogenase / Q000378:metabolism; D014461:Ultracentrifugation / Q000379:methods
D013385:Succinate Dehydrogenase