A convenient and sensitive fluorescence assay for phospholipid vesicles using diphenylhexatriene

✍ Scribed by E. London; G.W. Feigenson

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 481 KB
• Volume: 88
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90412-8

✦ Synopsis

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 pM DPH is proportional to the phospholipid concentration over a wide range. As little as 0.7 nmol (-0.5 pg 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.