TY - JOUR
T1 - A method for measuring anion transfer across membranes of hemoglobin-free cells and vesicles by continuous monitoring of fluorescence
AU - Darmon, A.
AU - Eidelman, O.
AU - Cabantchik, Z. I.
PY - 1982/1/15
Y1 - 1982/1/15
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0020482560&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(82)90591-7
DO - 10.1016/0003-2697(82)90591-7
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C2 - 7041697
AN - SCOPUS:0020482560
SN - 0003-2697
VL - 119
SP - 313
EP - 321
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -