Cohen C M, Kalish D I, Jacobson B S, Branton D
J Cell Biol. 1977 Oct;75(1):119-34. doi: 10.1083/jcb.75.1.119.
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.
将细胞与聚赖氨酸包被的聚丙烯酰胺珠结合后,已纯化了HeLa细胞质膜。在25℃、pH 5.0的蔗糖 - 醋酸盐缓冲液中,细胞与珠的附着及膜回收达到最大值。对哇巴因敏感的钠钾 - 三磷酸腺苷酶、膜结合的125I - 麦胚凝集素的测量以及化学分析表明,珠上的膜与通过传统方法分离的膜具有相当或更高的纯度。由于分离过程快速(约2.5小时),且产生的膜其原生质表面完全暴露,它应是标准分离技术的有用补充。