Purification of non-synaptic and synaptic mitochondria and plasma membranes from rat brain by a rapid Percoll gradient procedure

J Neurosci Methods. 1985 Jun;14(1):41-51. doi: 10.1016/0165-0270(85)90113-x.

Abstract

A method is described for the isolation and purification of non-synaptic and synaptic mitochondria and plasma membranes. The procedure involved differential centrifugation and successively a Ficoll gradient followed by a Percoll one. The method is rapid and maintains constant osmotic conditions. The Percoll-purified fractions show a lower density than those prepared by the conventional methods. The fractions were identified and characterized by enzyme markers and electron microscopy. The method will be useful for routine preparations rendering a good degree of homogeneity and purity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid Phosphatase / analysis
  • Animals
  • Brain Chemistry
  • Cell Fractionation / methods*
  • Cell Membrane / enzymology
  • Centrifugation, Density Gradient / methods
  • Mitochondria / enzymology*
  • Mitochondria / metabolism
  • NADPH-Ferrihemoprotein Reductase / analysis
  • Povidone
  • Rats
  • Rats, Inbred Strains
  • Silicon Dioxide
  • Sodium-Potassium-Exchanging ATPase / analysis
  • Subcellular Fractions / ultrastructure
  • Synaptosomes / enzymology*
  • Synaptosomes / metabolism

Substances

  • Percoll
  • Silicon Dioxide
  • NADPH-Ferrihemoprotein Reductase
  • Acid Phosphatase
  • Sodium-Potassium-Exchanging ATPase
  • Povidone