title: Isolation of canine neutrophil plasma membranes.
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 +/- 22nm), and small (119 +/- 11nm). 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.
authors.forename: R T
authors.initials: RT
authors.lastname: O'Donnell
authors.forename: B R
authors.initials: BR
authors.lastname: Andersen
mesh_terms: D015720:5'-Nucleotidase; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D002498:Centrifugation; D002850:Chromatography, Gel; D004285:Dogs; D006874:Hydrostatic Pressure; D009504:Neutrophils / Q000648:ultrastructure*; D009708:Nucleotidases / Q000378:metabolism
chemical_list: D009708:Nucleotidases; D015720:5'-Nucleotidase; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase