Methods in molecular biology (Clifton, N.J.), 2008, Vol.432, p.51
2008
Volltextzugriff (PDF)
Details
- Autor(en) / Beteiligte
- Titel
- Purification of Saccharomyces cerevisiae mitochondria by zone electrophoresis in a free flow device
- Ist Teil von
- Methods in molecular biology (Clifton, N.J.), 2008, Vol.432, p.51
- Ort / Verlag
- United States
- Erscheinungsjahr
- 2008
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1064-3745DOI: 10.1007/978-1-59745-028-7_4Titel-ID: cdi_pubmed_primary_18370010
- Format
- –
- Schlagworte
- Adenosine Triphosphatases - isolation & purification, Animals, Carrier Proteins - isolation & purification, Centrifugation - methods, Electron Transport Complex III - isolation & purification, Electron Transport Complex IV - isolation & purification, Horses, Immunoblotting, Membrane Proteins - isolation & purification, Mitochondria - chemistry, Mitochondria - ultrastructure, Mutation, Oxidative Phosphorylation, Saccharomyces cerevisiae - enzymology, Saccharomyces cerevisiae - genetics, Saccharomyces cerevisiae - ultrastructure, Saccharomyces cerevisiae Proteins - isolation & purification, Ubiquinone - analogs & derivatives