Isolation of cell plasma membranes on microcarrier culture beads.
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.
D000818:Animals; D002458:Cell Fractionation; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure*; D003238:Connective Tissue; D003911:Dextrans; D006867:Hydrolases / Q000032:analysis; D051379:Mice; D013724:Teratoma
D003911:Dextrans; C025614:sephadex; D006867:Hydrolases