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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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Anmerkungen
Besondere Publikation
Auf Hompepage verbergern
Publikationstyp
Artikel: Sammelbandbeitrag/Buchkapitel
Herausgeber
Posch, A.*
Schlagwörter
rat liver mitochondria; free flow electrophoresis; ysosomes; zone electrophoresis
Sprache
englisch
Veröffentlichungsjahr
2008
HGF-Berichtsjahr
2008
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Bandtitel
2D Page: Sample Preparation and Fractionation
Zeitschrift
Methods in Molecular Biology
Quellenangaben
Band: 424,
Seiten: 333-348
Verlag
Springer
Verlagsort
Berlin [u.a.]
Begutachtungsstatus
Peer reviewed
Institut(e)
Institute of Molecular Toxicology and Pharmacology (TOX)
Institute of Pathology (PATH)
Research Unit Analytical Pathology (AAP)
Institute of Pathology (PATH)
Research Unit Analytical Pathology (AAP)
POF Topic(s)
30203 - Molecular Targets and Therapies
30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
30205 - Bioengineering and Digital Health
30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
30205 - Bioengineering and Digital Health
Forschungsfeld(er)
Enabling and Novel Technologies
PSP-Element(e)
G-505200-001
G-500300-001
G-500390-001
G-500300-001
G-500390-001
Erfassungsdatum
2008-09-16