Quantitative isolation of liver mitochondria by zonal centrifugation |
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Authors: | Margaret A. Wilson Joseph Cascarano Wendy L. Wooten Cecil B. Pickett |
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Affiliation: | Department of Biology, University of California, Los Angeles, California 90024 USA |
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Abstract: | A method was developed using zonal centrifugation to recover liver mitochondria quantiatively and free of other cellular components from a sample of whole homogenate. The fractions containing mitochondria were identified by the distribution of cytochrome oxidase and these fractions contained over 90% of the total cytochrome oxidase recovered. The mitochondrial fractions were found to be only slightly contaminated by 5′-nucleotidase (plasma membranes), acid phosphatase (lysosomes), glucose-6-phosphatase (microsomes), and catalase (peroxisomes). There was no detectable contamination by nuclear DNA (nuclei). This method was used to quantitate total liver mitochondrial protein. The development of this procedure provides a means for following total changes in mitochondrial components during mitochondrial biogenesis. |
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