Keenan T W, Valivullah H M, Dunlevy J T
Department of Biochemistry and Nutrition, Virginia Polytechnic Institute, Blacksburg 24061.
Anal Biochem. 1989 Feb 15;177(1):194-8. doi: 10.1016/0003-2697(89)90039-0.
An aqueous polymer two phase partition method was adapted for isolation of plasma membranes from mammary gland of lactating rats. Plasma membranes isolated by this method were enriched 23-fold in activity of the plasma membrane marker phosphodiesterase I and were depleted, relative to homogenates, in activities of enzymes which are markers for intracellular organelles and endomembranes. Yields of plasma membranes obtained by this method were 0.3 +/- 0.1 mg protein/g wet tissue weight. Plasma membranes were isolated more rapidly and yields were more consistent than those obtained with density gradient centrifugation methods which have been applied for isolation of plasma membranes from mammary gland.
采用水相聚合物双相分配法从泌乳大鼠乳腺中分离质膜。用该方法分离的质膜中质膜标记物磷酸二酯酶I的活性提高了23倍,相对于匀浆,细胞内细胞器和内膜标记酶的活性降低。该方法获得的质膜产量为0.3±0.1mg蛋白质/克湿组织重量。与用于从乳腺中分离质膜的密度梯度离心法相比,该方法分离质膜的速度更快,产量更稳定。