A method for measuring anion transfer across red cell membranes by continuous monitoring of fluorescence
✍ Scribed by O. Eidelman; Z.I. Cabantchik
- Publisher
- Elsevier Science
- Year
- 1980
- Tongue
- English
- Weight
- 509 KB
- Volume
- 106
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A fluorometric method for continuous monitoring of anion transport across red blood cell membranes was developed. The method was based on the use of a fluorescent substrate, N-(2-aminoethylsulfonate)-7-nitrobenz-2-oxa-3-diazole) (NBD-taurine), whose intracellular fluorescence was quenched by hemoglobin. Cells loaded with NBD-taurine, free of extracellular probe, were suspended in various media, and the eftlux of probe was recorded as a time-dependent increase in fluorescence. The method was highly reproducible and was sensitive down to lo3 cells as well as to transport rates in the range lo-30 s. A single fluorescence trace allowed determination of the Michaelis-Menten parameters of NRD-taurine efflux.