critical comparison between two classical and a kit-based method for mitochondria isolation

• Published in: Proteomics (Weinheim) Vol. 9; no. 11; pp. 3209 - 3214
• Main Authors: Hartwig, Sonja, Feckler, Christian, Lehr, Stefan, Wallbrecht, Katrin, Wolgast, Heike, Müller-Wieland, Dirk, Kotzka, Jörg
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley-VCH Verlag 01-06-2009; WILEY-VCH Verlag; WILEY‐VCH Verlag; Wiley-VCH
• Subjects: 2-D gels; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Blotting, Western; Cell Fractionation - methods; Electrophoresis - methods; Electrophoresis, Gel, Two-Dimensional; Fundamental and applied biological sciences. Psychology; Mice; Miscellaneous; Mitochondria; Mitochondria, Liver; Proteins; Proteomics; Proteomics methods; Reproducibility of Results; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Subcellular fractionation; Succinate Dehydrogenase - metabolism; Ultracentrifugation - methods; Mitochondria; Subcellular fractionation; Proteomics; Isolation; Proteomics methods; Method; 2-D gels
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200800344

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.