Preparative scale isolation of basal-lateral plasma membranes from rat intestinal epithelial cells

Membrane Biochemistry
A K MircheffE M Wright

Abstract

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.

References

Apr 1, 1977·The Journal of Cell Biology·C H Van OsE M Wright
Apr 13, 1971·Biochimica Et Biophysica Acta·M FujitaM Nakao
Apr 1, 1969·Biochimica Et Biophysica Acta·J P Quigley, G S Gotterer
Jan 1, 1964·Analytical Biochemistry·A Dahlqvist

Related Concepts

Na(+)-K(+)-Exchanging ATPase
Cell Fractionation
Plasma Membrane
Centrifugation, Density Gradient
Mesothelium
Structure of Intestinal Gland
Intestines, Small
Microvilli
Succinate Dehydrogenase
Mutansucrase

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