A simple and fast method for the isolation of basolateral plasma membranes from rat small-intestinal epithelial cells.
A method was developed for the analytical and preparative isolation of basolateral plasma membranes from rat small intestine. They were separated on a self-orientating Percoll (modified colloidal silica) gradient starting with a heavy microsomal-membrane fraction and involving centrifugation at 48,000 g for 1 h. (Na+ + K+)-stimulated ATPase activity, used as a marker enzyme for the basolateral plasma membrane, is enriched 20-fold compared with that found in the homogenate of isolated intestinal epithelial cells.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D004848:Epithelium / Q000201:enzymology / Q000648:ultrastructure; D007421:Intestine, Small / Q000201:enzymology / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D011205:Povidone; D051381:Rats; D012822:Silicon Dioxide
C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone