A simple method for the isolation of microvilli from kidney brush border is described. The method depends on the preferential aggregation of other subcellular structures by bivalent metal ions. MgCl2 is added to a homogenate of cortical tissue prepared from frozen rabbit kidneys. Aggregated material is removed by a low-speed centrifugation and the supernatant centrifuged at 15000g to yield a pellet enriched in microvilli. This is resuspended and given a second treatment with Mg2+. The purified preparation is obtained after four short differential centrifugations. The six brush-border enzymes that were monitored were enriched 11–17-fold compared with the original homogenate and were obtained in about 10% yield. Marker enzymes for other subcellular components showed the preparation to be essentially free of mitochondria and to be less contaminated with endoplasmic reticulum and baso-lateral plasma membranes than are conventional brush-border preparations. The main contamination was of lysosomal origin, about half of which was attributable to adsorbed acid hydrolases rather than to intact lysosomes. The aggregated components in the low-speed pellet bound less Mg2+than did the microvillus fraction. A possible mechanism for the role of Mg2+is discussed.
Skip Nav Destination
Follow us on Twitter @Biochem_Journal
Article navigation
September 1974
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkAdvertising
Research Article|
September 01 1974
A rapid method for the preparation of microvilli from rabbit kidney Available to Purchase
Andrew G. Booth;
Andrew G. Booth
1Department of Biochemistry, University of Leeds, 9 Hyde Terrace, Leeds LS2 9LS, U.K.
Search for other works by this author on:
A. John Kenny
A. John Kenny
1Department of Biochemistry, University of Leeds, 9 Hyde Terrace, Leeds LS2 9LS, U.K.
Search for other works by this author on:
Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1974 London: The Biochemical Society
1974
Biochem J (1974) 142 (3): 575–581.
Citation
Andrew G. Booth, A. John Kenny; A rapid method for the preparation of microvilli from rabbit kidney. Biochem J 1 September 1974; 142 (3): 575–581. doi: https://doi.org/10.1042/bj1420575
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.
Biochemical Society Member Sign in
Sign InSign in via your Institution
Sign in via your InstitutionGet Access To This Article
Cited By
Follow us on Twitter @Biochem_Journal
Open Access for all
We offer compliant routes for all authors from 2025. With library support, there will be no author nor reader charges in 5 journals. Check here |
![]() View past webinars > |