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. 1977 Nov 15;168(2):187-94.
doi: 10.1042/bj1680187.

Rapid isolation of plasma membranes in high yield from cultured fibroblasts

D Thom, A J Powell, C W Lloyd, D A Rees

Rapid isolation of plasma membranes in high yield from cultured fibroblasts

D Thom et al. Biochem J. .

Abstract

1. A method was developed which allows the rapid preparation of pure plasma membranes in high yield from cultured fibroblasts. 2. Cells are lysed in hypo-osmotic borate/EDTA and, after differential centrifugation, the membranes collected by centrifugation on a sucrose barrier. 3. Electron microscopy of the isolated material shows large membrane vesicles essentially free from contaminating organelles. 4. There is no detectable activity of the endoplasmic-reticulum enzyme marker, NADH2--lipoamide oxidoreductase (EC 1.6.4.3), and that of succinate dehydrogenase (EC 1.3.99.1), a marker for mitochondria, is substantially decreased. Chemical compositions are in good agreement with previous observations. 5. This study confirms the usefulness of applied isotopic markers for isolating plasma membranes.

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