A simple and fast method for the isolation of basolateral plasma membranes from rat small-intestinal epithelial cells.

The Biochemical journal ·Vol. 186 ·No. 1 ·1980-01-15 ·Pages 177-81

Scalera V, Storelli C, Storelli-Joss C, Haase W, Murer H

Abstract

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.

MeSH Terms

Animals Cell Fractionation/methods Cell Membrane/enzymology,ultrastructure Centrifugation, Density Gradient Epithelium/enzymology,ultrastructure Intestine, Small/enzymology,ultrastructure Male Microscopy, Electron Povidone Rats Silicon Dioxide

Chemicals

Percoll Silicon Dioxide Povidone

Authors & Affiliations

5 authors, click to expand affiliations / ORCID

Scalera V

Storelli C

Storelli-Joss C

Haase W

Murer H

References (11)

11 references, click to expand

An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study.
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Distribution of (Na+-K+)-stimulated ATPase activity in rat intestinal mucosa.
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Further biochemical and morphological studies of granule fractions from rabbit heterophil leukocytes.
J Cell Biol. 1970 Jun;45(3):586-97 PMID: 5459943
Differential isolation of microvillous and basolateral plasma membranes from intestinal mucosa: mutually exclusive distribution of digestive enzymes and ouabain-sensitive ATPase.
Biochim Biophys Acta. 1972 Aug 9;274(2):336-47 PMID: 4340259
Preparation and characterization of the lateral and basal plasma membranes of the rat intestinal epithelial cell.
Biochem J. 1972 Aug;128(5):1329-38 PMID: 4566193
Glucose transport in isolated brush-border and lateral-basal plasma-membrane vesicles from intestinal epithelial cells.
Biochim Biophys Acta. 1974 Apr 29;345(2):170-9 PMID: 4366460
Basolateral plasma membranes of intestinal epithelial cells. Identification by lactoperoxidase-catalysed iodination and isolation after density perturbation with digitonin.
Biochem J. 1975 Oct;152(1):71-84 PMID: 129058
Transport of p-aminohippuric acid by plasma membrane vesicles isolated from rat kidney cortex.
Pflugers Arch. 1976 Feb 24;361(3):269-77 PMID: 943770
The surface membrane of the small intestinal epithelial cell. I. Localization of adenyl cyclase.
Biochim Biophys Acta. 1976 May 21;433(3):509-19 PMID: 179591
Analytical isolation of plasma membranes of intestinal epithelial cells: identification of Na, K-ATPase rich membranes and the distribution of enzyme activities.
J Membr Biol. 1976 Sep 17;28(4):309-33 PMID: 136516
Subcellular distribution of nucleotide cyclases in rat intestinal epithelium.
Am J Physiol. 1978 Nov;235(5):E539-45 PMID: 31794

Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj1860177

Published

1980-01-15

Pages

177-81

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1161517

Subset

External Links

PubMed source DOI: 10.1042/bj1860177 PMC Full Text View journal details

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A simple and fast method for the isolation of basolateral plasma membranes from rat small-intestinal epithelial cells.

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