Preparation of plasma membrane from isolated neurons.
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
D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology; D002466:Cell Nucleolus; D002467:Cell Nucleus; D002499:Centrifugation, Density Gradient; D002540:Cerebral Cortex / Q000166:cytology / Q000201:enzymology; D003580:Cytochromes; D003576:Electron Transport Complex IV / Q000032:analysis; D005958:Glucosephosphates / Q000032:analysis; D008722:Methods; D008854:Microscopy, Electron; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D008995:Monoamine Oxidase / Q000032:analysis; D009249:NADP; D009474:Neurons / Q000166:cytology* / Q000201:enzymology; D010088:Oxidoreductases / Q000032:analysis; D011817:Rabbits; D013395:Sucrose
D003580:Cytochromes; D005958:Glucosephosphates; D009249:NADP; D013395:Sucrose; D010088:Oxidoreductases; D008995:Monoamine Oxidase; D003576:Electron Transport Complex IV; D000251:Adenosine Triphosphatases