Tissue-culture Fractionation. Zonal Centrifugation of Lettrée Cells Homogenized by Glycerol-induced Lysis: Subfractionation of Membranes in Sucrose Gradients

Overview

Journal Biochem J

Specialty Biochemistry

Date 1979 Jul 15

PMID 496904

Citations 1

Authors

J M Graham

K H Coffey

Affiliations

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Abstract

1. Lettrée cells were grown intraperitoneally in MF-1 mice. 2. Cells that were loaded with glycerol were swollen in 0.1 M-sucrose and disrupted by Dounce homogenization. 3. Early-passage Lettrée cells were more easily disrupted than late-passage cells by this method, and the former produced larger fragments of plasma membrane. 4. The membranes were fractionated initially in sucrose gradients (on the basis of sedimentation rate) in a BXIV zonal rotor. 5. Fractions from this gradient were further resolved in isopycnic sucrose gradients. 6. Plasma-membrane and endoplasmic-reticulum fractions were recovered in good yield and high purity.

Citing Articles

Tissue-culture cell fractionation. Fractionation of cellular membranes from 125I/lactoperoxidase-labelled Lettrée cells homogenized by bicarbonate-induced lysis: resolution of membranes by zonal centrifugation and in sucrose and metrizamide....

Graham J, Coffey K Biochem J. 1979; 182(1):173-80.

PMID: 496905 PMC: 1161246. DOI: 10.1042/bj1820173.

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