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

doi:10.1016/0005-2736(89)90382-9

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

School of Medicine

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

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 language	English (US)
Pages (from-to)	77-81
Number of pages	5
Journal	BBA - Biomembranes
Volume	983
Issue number	1
DOIs	https://doi.org/10.1016/0005-2736(89)90382-9
State	Published - Jul 24 1989

Keywords

(Dog kidney cortex)
Basolateral membrane
Brush-border membrane

ASJC Scopus subject areas

Biophysics
Biochemistry
Cell Biology

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10.1016/0005-2736(89)90382-9

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@article{f121538fa91b42e58fc7413596ffc206,

title = "Techniques for isolation of brush-border and basolateral membrane vesicles from dog kidney cortex",

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.",

keywords = "(Dog kidney cortex), Basolateral membrane, Brush-border membrane",

author = "Hilden, {Shirley A.} and Johns, {Conrado A.} and Guggino, {William B.} and Madias, {Nicolaos E.}",

note = "Funding Information: This work was supportedin part bY National In-stituteso f Health grants DK-32752( to W.B.G.) and DK-38155( to N.E.M.) and a grantf romD ialysisC linic, Inc. (to S.A.H.). Copyright: Copyright 2014 Elsevier B.V., All rights reserved.",

year = "1989",

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doi = "10.1016/0005-2736(89)90382-9",

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AU - Hilden, Shirley A.

AU - Johns, Conrado A.

AU - Guggino, William B.

AU - Madias, Nicolaos E.

N1 - Funding Information: This work was supportedin part bY National In-stituteso f Health grants DK-32752( to W.B.G.) and DK-38155( to N.E.M.) and a grantf romD ialysisC linic, Inc. (to S.A.H.). Copyright: Copyright 2014 Elsevier B.V., All rights reserved.

PY - 1989/7/24

Y1 - 1989/7/24

N2 - 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.

AB - 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.

KW - (Dog kidney cortex)

KW - Basolateral membrane

KW - Brush-border membrane

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

Abstract

Keywords

ASJC Scopus subject areas

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