Techniques for isolation of brush-border and basolateral membrane vesicles from dog kidney cortex
SA Hilden, CA Johns, WB Guggino… - Biochimica et Biophysica …, 1989 - Elsevier
SA Hilden, CA Johns, WB Guggino, NE Madias
Biochimica et Biophysica Acta (BBA)-Biomembranes, 1989•ElsevierTwo 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 Mg 2+. The Mg 2+ treatment produced a
pellet (crude BLMV) which was added to Percoll and centrifuged to produce purified BLMV.
The supernatant after Mg 2+ treatment eventually yielded pure BBMV after additional Mg 2+ …
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 Mg 2+. The Mg 2+ treatment produced a
pellet (crude BLMV) which was added to Percoll and centrifuged to produce purified BLMV.
The supernatant after Mg 2+ treatment eventually yielded pure BBMV after additional Mg 2+ …
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.
Elsevier