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PMID: 21192 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells.

The Journal of cell biology ·Vol. 75 ·No. 1 ·1977-10-00 ·Pages 119-34

Cohen CM, Kalish DI, Jacobson BS, Branton D

Abstract

HeLa cell plasma membranes have been purified after binding cells to polylysine-coated polyacrylamide beads. Cell attachment to beads and membrane recovery were maximal in a sucrose-acetate buffer, pH 5.0, at 25 degrees C. Measurements of ouabain-sensitive NaK-adenosine triphosphatase, membrane-bound 125I-wheat germ agglutinin, and chemical analyses showed that membranes on beads were of comparable or greater purity than membranes isolated by conventional methods. Because the isolation procedure is rapid (approximately 2.5 h), and produces membranes whose protoplasmic surfaces are fully exposed, it should be a useful supplement to standard isolation techniques.

MeSH Terms
Cell Count Cell Fractionation/methods Cell Membrane/analysis,ultrastructure DNA HeLa Cells Hydrogen-Ion Concentration Membrane Lipids/analysis Membrane Proteins/analysis Osmolar Concentration Polylysine Temperature
Chemicals
Membrane Lipids Membrane Proteins Polylysine DNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Cohen C M
Kalish D I
Jacobson B S
Branton D
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26 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1977-10-00
Pages
119-34
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2111560
Subset
IM
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