A method for measuring anion transfer across membranes of hemoglobin-free cells and vesicles by continuous monitoring of fluorescence

A. Darmon, O. Eidelman, Z. I. Cabantchik*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

A fluorimetric method for fast and continuous monitoring of anion transport across membrane vesicles was developed. The method was based on the use of the fluorescent substrate N-(2-aminoethyl-sulfonate)-7-nitrobenz-2-oxa-3-diazole (NBD-taurine), whose fluorescence was quenched by anti-NBD antibodies added to the external flux medium. Various types of vesicles loaded with NBD-taurine and free of extravesicular fluorescent substrate were suspended in antibody-containing flux medium, and the time-dependent egress of probe was recorded continuously as an exponential decrease in intravesicular fluorescence. The new method circumvents the need for separating vesicles from medium. It displays high sensitivity, requires small quantities of vesicles, and gives reproducible transport profiles of transport rates down to the 0.2-min range.

Original languageEnglish
Pages (from-to)313-321
Number of pages9
JournalAnalytical Biochemistry
Volume119
Issue number2
DOIs
StatePublished - 15 Jan 1982

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