Plasma membrane isolation on DEAE-Sephadex beads.
A much-simplified method for the purification of plasma membranes of cultured cells is presented, based upon the attachment of viable cells to nitrocellulose-treated DEAE-Sephadex beads, and their subsequent shearing by hypotonic lysis, agitation on a vortex mixer and sonication. The method is suggested by an older procedure involving attachment to poly-(L-lysine)-coated glass or polyacrylamide beads; the preparation involved in the present method, however, is considerably easier, more rapid and less expensive. Recovery of L-cell plasma membrane marker enzyme activities is approx. 25%, while contamination by internal membrane markers is much less than 1%.
D000818:Animals; D002417:Cattle; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism*; D002478:Cells, Cultured; D002642:Chick Embryo; D002852:Chromatography, Ion Exchange; D003238:Connective Tissue / Q000201:enzymology*; D051379:Mice