title: Optimal isolation of mitochondria for proteomic analyses.
abstract: Considering the key role of mitochondria in cellular (dys)functions, we compared a standard isolation protocol, followed by lysis in urea/detergent buffer, with a commercially available isolation buffer that rapidly yields a mitochondrial protein fraction. The standard protocol yielded significantly better overall resolution and coverage of both the soluble and membrane mitochondrial proteomes; although the kit was faster, it resulted in recovery of only approximately 56% of the detectable proteome. The quality of "omic" analysis depends on sample handling; for large-scale protein studies, well-resolved proteomes are highly dependent on the purity of starting material and the rigor of the extraction protocol.
authors.affiliation: Neuro-Cell Biology Laboratory, School of Science and Health, University of Western Sydney, Campbelltown, NSW 2560, Australia; Molecular Medicine Research Group, School of Medicine, University of Western Sydney, Campbelltown, NSW 2560, Australia.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Stimpson
authors.affiliation: Molecular Medicine Research Group, School of Medicine, University of Western Sydney, Campbelltown, NSW 2560, Australia; Molecular Physiology Unit, School of Medicine, University of Western Sydney, Campbelltown, NSW 2560, Australia. Electronic address: j.coorssen@uws.edu.au.
authors.forename: Jens R
authors.initials: JR
authors.lastname: Coorssen
authors.affiliation: Neuro-Cell Biology Laboratory, School of Science and Health, University of Western Sydney, Campbelltown, NSW 2560, Australia; Molecular Medicine Research Group, School of Medicine, University of Western Sydney, Campbelltown, NSW 2560, Australia. Electronic address: s.myers@uws.edu.au.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Myers
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D006801:Humans; D008214:Lymphocytes; D008928:Mitochondria; D040901:Proteomics / Q000379:methods*
keywords: Mitochondria; Proteomics; Two-dimensional gel electrophoresis