Techniques for isolation of brush-border and basolateral membrane vesicles from dog kidney cortex

Shirley A. Hilden, Conrado A. Johns, William B. Guggino, Nicolaos E. Madias

Research output: Contribution to journalArticlepeer-review

Abstract

Two methods are reported for renal membrane preparation from the dog kidney cortex. One method is a simultaneous preparation of brush-border (BBMV) and basolateral (BLMV) membranes. Using readily available laboratory equipment, differential centrifugation produced a supernatant which was treated with Mg2+. The Mg2+ treatment produced a pellet (crude BLMV) which was added to Percoll and centrifuged to produce purified BLMV. The supernatant after Mg2+ treatment eventually yielded pure BBMV after additional Mg2+ precipitations. The second method used an acidic medium in conjunction with divalent-cation precipitation to prepare BBMV. Whichever method was used, BBMV and BLMV showed appropriate enzyme and transport activities.

Original languageEnglish (US)
Pages (from-to)77-81
Number of pages5
JournalBBA - Biomembranes
Volume983
Issue number1
DOIs
StatePublished - Jul 24 1989

Keywords

  • (Dog kidney cortex)
  • Basolateral membrane
  • Brush-border membrane

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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