Isolation of canine neutrophil plasma membranes

Robert T O'Donnell, B. R. Andersen

Research output: Contribution to journalArticle

Abstract

This paper describes the isolation of plasma membrane vesicles formed by nitrogen cavitation of canine neutrophils. Plasma membranes from disrupted cells were separated from other membranes and organelles by Percoll-density gradient centrifugation. Transmission electron microscopic examination of membrane preparations chromatographed on either Sephacryl S-1000 or Sepharose 4B revealed that two populations of plasma membrane vesicles were formed: large (176 ± 22 nm), and small (119 ± 11 nm). Purified large vesicles were separated from Percoll and contaminating cytosol by Sephacryl S-1000 chromatography. Small vesicles were obtained free of Percoll by recavitating purified large vesicles. Problems encountered due to the presence of a soluble 5'-nucleotidase inhibitor also are discussed. Large and small membrane vesicles were separated into adherent and non-adherent populations by affinity chromatography on either concanavalin A-sepharose or lentil lectin-Sepharose columns.

Original languageEnglish (US)
Pages (from-to)353-368
Number of pages16
JournalJournal of Leukocyte Biology
Volume38
Issue number3
StatePublished - 1985
Externally publishedYes

ASJC Scopus subject areas

  • Cell Biology

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