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  • Title: Improved method for the isolation of rat liver plasma membrane.
    Author: Brown AE, Lok MP, Elovson J.
    Journal: Biochim Biophys Acta; 1976 Mar 19; 426(3):418-32. PubMed ID: 817740.
    Abstract:
    An improved method for the isolation of plasma membrane from rat liver is presented. Gentle homogenization of perfused livers in buffered isotonic KCL, followed by direct flotation of a low-speed nuclear pellet through a discontinuous sucrose density gradient results in a 32% yield, and 25-fold enrichment for the plasma membrane marker, phosphodiesterase I, in a crude plasma membrane fraction. This fraction contains less than 1% of the mitochondria, and endoplasmic reticulum present in the original homogenate, but is more heavily contaminated with lysosomes and Golgi membrane. Vigorous mechanical disruption of this material, followed by a second discontinuous sucrose density gradient, gives a light plasma membrane fraction with an 80-fold purification and 20% yield of phosphodiesterase I over the original homogete (with further reduction of contaminants).
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