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Plasma membrane isolation on DEAE-Sephadex beads
Authors:Louis J. Gotlib  David B. Searls
Affiliation:Department of Biology, The Johns Hopkins University, 34th and N. Charles Streets, Baltimore, MD 21218 U.S.A.
Abstract: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%.
Keywords:Plasma membrane isolation  Adherence  Glass bead  Cell surface  (DEAE-Sephadex)
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