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Isolation of mitochondria from cultured cells and liver tissue biopsies for molecular and biochemical analyses.

Methods Mol. Biol. 1295, 87-97 (2015)
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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
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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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
ISSN (print) / ISBN 1064-3745
e-ISSN 1940-6029
Konferenztitel Proteomic Profiling : Methods and Protocols
Quellenangaben Band: 1295, Heft: , Seiten: 87-97 Artikelnummer: , Supplement: ,
Verlag Springer
Verlagsort Berlin [u.a.]
Begutachtungsstatus Peer reviewed