The lucigenin assay: measuring anion transport in lipid vesicles

14 June 2023, Version 1
This content is a preprint and has not undergone peer review at the time of posting.


Synthetic anion transporters are developed to transport anions across lipid membranes with the long-term perspective of biological applications. The lucigenin assay is a popular tool to study their transport of chloride and other anions in liposomes. It relies on the quenching of the fluorescence of encapsulated lucigenin by anions, which can be monitored by fluorescence spectroscopy. This article provides a tutorial introduction to the practical use and understanding of the lucigenin assay. It describes in detail how to use this assay to monitor chloride/nitrate antiport in liposomes, process and interpret the data, and solve common issues. Variations of the assay enabling the investigation of the transport of other anions and transport mechanisms are discussed. Furthermore, a zwitterionic analogue of lucigenin is demonstrated to have advantages for use in specific cases.


Anion transport
Fluorescent probes
Fluorescence spectroscopy
Supramolecular Chemistry

Supplementary materials

Supporting information
Stern-Volmer fluorescence quenching plots for lucigenin and SPBA, structures of compounds, DLS data, and additional transport data.


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