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A convenient and sensitive fluorescence assay for phospholipid vesicles using diphenylhexatriene

Authors
Journal
Analytical Biochemistry
0003-2697
Publisher
Elsevier
Publication Date
Volume
88
Issue
1
Identifiers
DOI: 10.1016/0003-2697(78)90412-8

Abstract

Abstract When phospholipid vesicles are added to an aqueous solution of 1,6-diphenyl-1,3,5-hexatriene (DPH) a fluorescence enhancement of up to several hundredfold is observed which can be used for a determination of phospholipid concentration. Fluorescence enhancement of 2 μ m DPH is proportional to the phospholipid concentration over a wide range. As little as 0.7 nmol (∼0.5 μg of phospholipid) can be determined to within ±10%. The fluorescence is a function of the type of phospholipid used, salt concentration, and time of incubation. Protein and detergents also enhance DPH fluorescence but to a much smaller extent. Optimal conditions for the assay are presented. Use of this assay to detect phospholipid vesicles fractionated by size on a Sepharose 4B column is illustrated. In this case the method compares favorably to more classical methods of analysis in terms of sensitivity, accuracy, and time involved.

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