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J. Biophys. and Biochem. Cytol., Vol 8, 413-422, Copyright © 1960 by Rockefeller University Press

ARTICLE

THE ISOLATION OF A CELL MEMBRANE FRACTION FROM RAT LIVER

David M. Neville Jr. M.D.1

1 From the Department of Pathology, University of Rochester School of Medicine and Dentistry, Rochester, New York.

Dr. Neville's present address is National Institute of Mental Health, National Institutes of Health, Bethesda, Maryland

A procedure is described for isolating cell membranes from rat liver homogenates. 20 gm. of rat liver was homogenized in a Dounce homogenizer in ice cold water buffered to pH 7.5 with NaHCO3, rupturing all of the cells and most nuclei. The diluted homogenate was filtered through cheesecloth to remove precipitated nucleoprotein and centrifuged at 1500 g, 10 minutes, to sediment a crude membrane fraction. The membrane containing sediment was recentrifuged 3 times in conical tubes (1220 g, 10 minutes), the top layer of the 2-layered sediment being retained. Flotation in a sucrose solution d = 1.22 freed the preparation from contaminating cell fragments and nuclear membranes not previously disintegrated. The floating material sim0.4 ml. was quite homogeneous and consisted of thin amorphous membranes. Electron micrographs revealed numerous double profiles similar in shape and dimensions to apposed liver cell membranes in intact tissue.

Submitted on February 15, 1960


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