title: Improved method for the isolation of rat liver plasma membrane.
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).
mesh_terms: D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient; D005260:Female; D006056:Golgi Apparatus / Q000648:ultrastructure; D008099:Liver / Q000201:enzymology / Q000648:ultrastructure*; D008854:Microscopy, Electron; D009251:NADPH-Ferrihemoprotein Reductase / Q000032:analysis; D051381:Rats
chemical_list: D009251:NADPH-Ferrihemoprotein Reductase; D000135:Acid Phosphatase