Preparation of plasma membrane from isolated neurons

J Cell Biol. 1972 Jun;53(3):654-61. doi: 10.1083/jcb.53.3.654.

Abstract

A bulk fraction enriched with respect to neuronal cell bodies was used as starting material for the isolation of neuronal plasma membrane The cells were gently homogenized in isotonic sucrose and a crude membrane containing fraction sedimented at 3000 g. Subsequently, the membrane fraction was purified on a discontinuous sucrose density gradient between 35% and 25 5% sucrose (w/w). Enzymatic analyses showed a 4-5-fold enrichment in plasma membrane markers, and a 10-15% contamination of mitochondrial and microsomal material. Electron micrographs of the membrane fraction confirmed the enzymatic data Fragmented membranes were found, mainly in vesicular form No ribosomes, but a few mitochondria and some multilamellar membranes were seen

MeSH terms

  • Adenosine Triphosphatases / analysis
  • Animals
  • Cell Fractionation
  • Cell Membrane* / enzymology
  • Cell Nucleolus
  • Cell Nucleus
  • Centrifugation, Density Gradient
  • Cerebral Cortex / cytology
  • Cerebral Cortex / enzymology
  • Cytochromes
  • Electron Transport Complex IV / analysis
  • Glucosephosphates / analysis
  • Methods
  • Microscopy, Electron
  • Microsomes / enzymology
  • Mitochondria / enzymology
  • Monoamine Oxidase / analysis
  • NADP
  • Neurons / cytology*
  • Neurons / enzymology
  • Oxidoreductases / analysis
  • Rabbits
  • Sucrose

Substances

  • Cytochromes
  • Glucosephosphates
  • NADP
  • Sucrose
  • Oxidoreductases
  • Monoamine Oxidase
  • Electron Transport Complex IV
  • Adenosine Triphosphatases