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Isolation of cell plasma membranes on microcarrier culture beads
Authors:Louis J Gotlib
Institution:Department of Biology, The Johns Hopkins University, 34th and N. Charles Streets, Baltimore, MD 21218 U.S.A.
Abstract:A simple, efficient method for the purification of plasma membranes from cultured cells is presented. Membrane purification is effected by attachment of viable cells to commercially available microcarrier culture beads, followed by lysis of the cells, agitation on a vortex mixer and sonication. Optimal conditions for each step of the procedure are described. Enzyme markers from plasma membranes are purified 10–20-fold relative to whole cell homogenates while internal membrane markers are depleted 10–20-fold.
Keywords:Plasma membrane isolation  Polycationic bead  FMH  fat head minnow cells
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