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