Preparative scale isolation of basal-lateral plasma membranes from rat intestinal epithelial cells.
A simple, efficient procedure is described for the preparative scale isolation of basal-lateral membranes from the rat intestinal epithelium. The intestinal mucosa was mildly homogenized and soluble protein and RNA were separated from the homogenate by differential centrifugation. The basal-lateral membranes were then separated from nuclei, mitochondria, and brush border membranes by differential centrifugation in a medium close to the equilibrium density of the basal-lateral membranes. Final purification of the basal-lateral membranes was achieved on a linear density gradient in a high-capacity zonal rotor. The final product (usually at least 40 mg protein) represented a 34% yield of basal-lateral membranes purified 18-fold with respect to protein, 26-fold with respect to brush border membranes, and 53-fold with respect to mitochondria.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient / Q000379:methods; D004848:Epithelium / Q000648:ultrastructure; D007413:Intestinal Mucosa / Q000648:ultrastructure*; D007421:Intestine, Small / Q000648:ultrastructure*; D008297:Male; D008871:Microvilli / Q000648:ultrastructure; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis; D013385:Succinate Dehydrogenase / Q000032:analysis; D013393:Sucrase / Q000032:analysis
D013385:Succinate Dehydrogenase; D013393:Sucrase; D000254:Sodium-Potassium-Exchanging ATPase