The isolation of synaptosomes and synaptic plasma membranes from bovine cerebral cortex
Date
1977
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Abstract
A method is described for the preparation of synaptosomes and synaptic plasma membranes from Bovine cerebral cortex. After homogenization of the tissue in isotonic sucrose, differential centrifugation yielded a crude mitochondrial fraction (P2) which was purified by centrifugation on a Ficoll-sucrose gradient. Assay of occluded Lactate Dehydrogenase and electron microscopy confirmed that synaptosomes survived the fractionation procedure and appeared in the P2B and P2C fractions, sedimenting to a density of approximately 1.117. The final membrane preparation had a specific 5' -nucleotidase activity (units/mg protein) over 10 times higher than the brain homogenate. Membrane fragments derived from outer mitochondrial membranes and microsomes appear to be the major contaminants. Gel electrophoresis revealed proteins migrating at approximately the same rate as contractile proteins.
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cows
veterinary medicine
research