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Open Access Green as soon as Postprint is submitted to ZB.
Isolation of functional mitochondria from cultured cells and mouse tissues.
Methods Mol. Biol. 1567, 15-32 (2017)
Mitochondria serve as the center stage for a number of cellular processes, including energy production, apoptosis, ion homeostasis, iron and copper processing, steroid metabolism, de novo pyrimidine, and heme biosynthesis. The study of mitochondrial function often requires the purification of intact and respiratory-competent organelles. Here, we provide detailed protocols to isolate functional mitochondria from various types of mammalian cells and mouse tissues, in both crude and pure forms. We introduce the use of nitrogen cavitation for the disruption of plasma membrane and the reproducible isolation of mitochondria-enriched fractions of high yield. Mitochondria that are isolated by these procedures are intact and coupled and can directly be used for several downstream analyses, such as measurements of oxygen consumption and calcium buffering capacity.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Crude Mitochondria ; Mitochondrial Integrity ; Nitrogen Cavitation ; Organelle Isolation ; Pure Mitochondria
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Conference Title
Mitochondria
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 1567,
Pages: 15-32
Publisher
Springer
Publishing Place
Berlin [u.a.]
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Diabetes and Obesity (IDO)