Techniques for isolation of brush-border and basolateral membrane vesicles from dog kidney cortex.
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.
Department of Medicine, Tufts University School of Medicine, Boston, MA.
S A
SA
Hilden
C A
CA
Johns
W B
WB
Guggino
N E
NE
Madias
D000818:Animals; D001692:Biological Transport; D002413:Cations, Divalent; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure*; D004285:Dogs; D004798:Enzymes / Q000032:analysis; D007672:Kidney Cortex / Q000378:metabolism / Q000648:ultrastructure*; D008871:Microvilli / Q000378:metabolism / Q000648:ultrastructure*; D014461:Ultracentrifugation / Q000379:methods
D002413:Cations, Divalent; D004798:Enzymes