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Open Access Green as soon as Postprint is submitted to ZB.
Purification of Saccharomyces cerevisiae mitochondria by zone electrophoresis in a free flow device.
Methods Mol. Biol. 432, 51-64 (2008)
This chapter describes the isolation of yeast mitochondria by differential centrifugation followed by mitochondrial purification through zone electrophoresis (ZE) using a free flow device (FFE). Starting from a yeast colony, cultures are grown under respiratory conditions to logarithmic phase. Cells are collected, their cell walls enzymatically disintegrated and the resulting spheroplasts are homogenized. Mitochondria are pre-fractionated from this homogenate by differential centrifugation. With the focus on further purification, pre-fractionated mitochondria are subjected to ZE-FFE. In ZE-FFE, mitochondria are transported with the buffer flow through the separation chamber and purified from contaminants by specific deflection through a perpendicularly oriented electric field. The purified mitochondria can be collected by centrifugation and used for further experiments and analysis such as sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting, 2-DE or electron microscopy.
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Publication type
Article: Journal article
Document type
Scientific Article
Editors
Pflieger, D.* ; Rossier, J.*
Keywords
Mitochondria; free flow electrophoresis; zone electrophoresis; Saccharomyces cerevisiae
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Conference Title
Organelle Proteomics
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 432,
Pages: 51-64
Publisher
Springer
Publishing Place
Berlin [u.a.]
Reviewing status
Peer reviewed