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Open Access Green as soon as Postprint is submitted to ZB.
Isolation of mitochondria from cultured cells and liver tissue biopsies for molecular and biochemical analyses.
Methods Mol. Biol. 1295, 87-97 (2015)
We recently reported a new method to isolate functionally intact mitochondria from cell culture and small tissue samples (Schmitt et al., Anal Biochem 443(1):66-74, 2013). This method comprises a semi-automated cell rupture, termed pump controlled cell rupture system (PCC), which can be precisely adjusted to the specific cellular source of isolation and which can be tightly controlled (Schmitt et al., Anal Biochem 443(1):66-74, 2013). Here we provide a detailed hands-on protocol of this PCC method which results in an efficient cell breakage but preserving the mitochondrial integrity. Upon subsequent purification steps, the obtained mitochondrial fraction meets the quality and purity required for molecular analyses, e.g. proteomic comparisons, as well as for biochemical analyses, e.g. determination of diverse enzymatic activities.
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Publication type
Article: Journal article
Document type
Scientific Article
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Conference Title
Proteomic Profiling : Methods and Protocols
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 1295,
Pages: 87-97
Publisher
Springer
Publishing Place
Berlin [u.a.]
Reviewing status
Peer reviewed
Institute(s)
Institute of Molecular Toxicology and Pharmacology (TOX)
Institute of Pathology (PATH)
Research Unit Analytical Pathology (AAP)
Molekulare Endokrinologie und Metabolismus (MEM)
Institute of Pathology (PATH)
Research Unit Analytical Pathology (AAP)
Molekulare Endokrinologie und Metabolismus (MEM)