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
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- Bernhard W., Avrameas S. Ultrastructural visualization of cellular carbohydrate components by means of concanavalin A. Exp Cell Res. 1971 Jan;64(1):232–236. doi: 10.1016/0014-4827(71)90217-5. [DOI] [PubMed] [Google Scholar]
- Blomstrand C., Hamberger A. Amino acid incorporation in vitro into protein of neuronal and glial cell-enriched fractions. J Neurochem. 1970 Aug;17(8):1187–1195. doi: 10.1111/j.1471-4159.1970.tb03367.x. [DOI] [PubMed] [Google Scholar]
- Blomstrand C., Hamberger A. Protein turnover in cell-enriched fractions from rabbit brain. J Neurochem. 1969 Sep;16(9):1401–1407. doi: 10.1111/j.1471-4159.1969.tb05992.x. [DOI] [PubMed] [Google Scholar]
- Dallner G., Siekevitz P., Palade G. E. Biogenesis of endoplasmic reticulum membranes. II. Synthesis of constitutive microsomal enzymes in developing rat hepatocyte. J Cell Biol. 1966 Jul;30(1):97–117. doi: 10.1083/jcb.30.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
- EMMELOT P., BOS C. J., BENEDETTI E. L., RUEMKE P. STUDIES ON PLASMA MEMBRANES. I. CHEMICAL COMPOSITION AND ENZYME CONTENT OF PLASMA MEMBRANES ISOLATED FROM RAT LIVER. Biochim Biophys Acta. 1964 Jul 15;90:126–145. doi: 10.1016/0304-4165(64)90125-4. [DOI] [PubMed] [Google Scholar]
- HUEBSCHER G., WEST G. R. SPECIFIC ASSAYS OF SOME PHOSPHATASES IN SUBCELLULAR FRACTIONS OF SMALL INTESTINAL MUCOSA. Nature. 1965 Feb 20;205:799–800. doi: 10.1038/205799a0. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lehninger A. L. The neuronal membrane. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1069–1080. doi: 10.1073/pnas.60.4.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lutz F., Frimmer M. A new method for the isolation of plasma mebranes from pig liver. A combined procedure using 50 Hz-vibration of a sucrose gradient. Hoppe Seylers Z Physiol Chem. 1970 Dec;351(12):1429–1434. doi: 10.1515/bchm2.1970.351.2.1429. [DOI] [PubMed] [Google Scholar]
- McKeel D. W., Jarett L. Preparation and characterization of a plasma membrane fraction from isolated fat cells. J Cell Biol. 1970 Feb;44(2):417–432. doi: 10.1083/jcb.44.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
- NEVILLE D. M., Jr The isolation of a cell membrane fraction from rat liver. J Biophys Biochem Cytol. 1960 Oct;8:413–422. doi: 10.1083/jcb.8.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Waldmin A. A., Alm G. Effects of sucrose on determination of DNA by the method of Kissane and Robins. Anal Biochem. 1970 Jul;36(1):247–251. doi: 10.1016/0003-2697(70)90357-x. [DOI] [PubMed] [Google Scholar]