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Open Access Green as soon as Postprint is submitted to ZB.
Isolation of highly pure rat liver mitochondria with the aid of zone-electrophoresis in a free flow device (ZE-FFE).
In: 2D Page: Sample Preparation and Fractionation. Berlin [u.a.]: Springer, 2008. 333-348 (Methods Mol. Biol. ; 424)
This protocol describes the purification of mitochondria from rat liver with the aid of zone electrophoresis in a free flow device (ZE-FFE). Starting from liver homogenate, cell debris and nuclei are removed by low speed centrifugation. A crude mitochondrial fraction is obtained by medium speed centrifugation and is further purified by washing followed by a Nycodenz® gradient centrifugation. Lysosomes and microsomes are located at the upper parts of the gradient, whereas mitochondria are found in the medium part of the gradient. A subsequent purification step with ZE-FFE efficiently removes remaining lysosomes and microsomes and, importantly, damaged mitochondrial structures. The resulting purified mitochondria can be concentrated by centrifugation and used for further experiments. Finally, possible modifications of this protocol with respect to the isolation of pure lysosomes are discussed.
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Publication type
Article: Edited volume or book chapter
Editors
Posch, A.*
Keywords
rat liver mitochondria; free flow electrophoresis; ysosomes; zone electrophoresis
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Book Volume Title
2D Page: Sample Preparation and Fractionation
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 424,
Pages: 333-348
Publisher
Springer
Publishing Place
Berlin [u.a.]
Non-patent literature
Publications
Reviewing status
Peer reviewed