A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:00Title
- 00:50Introduction
- 01:22Generation of 8-aminonaphthalene-1,3,6-trisulfonate (ANTS) Filled Liposomes
- 04:12Fluorescent Solution Preparation
- 05:02Fluorescence Instrument Setup
- 06:21Performing a Gramicidin-based Fluorescence Assay
- 07:34Data Analysis
- 09:16Representative Results
- 10:25Conclusion
We introduce a fast fluorescence-based assay that monitors the rate of fluorescence quenching as a measure of gramicidin channel activity. The gramicidin channels are used as molecular force transducers to monitor changes in lipid bilayer properties as sensed by bilayer spanning proteins.
Tags
Gramicidin-based Fluorescence AssaySmall MoleculesLipid Bilayer PropertiesAdsorptionAmphiphilesBilayer/solution InterfaceHydrophobic InteractionsMembrane ProteinsProtein Function ModulationOff-target Drug EffectsElectrophysiological AssayLinear Gramicidin ChannelsProbesMembrane EnvironmentFluorescence QuenchingFluorophore-loaded Large Unilamellar VesiclesQuencherFluorescence Indicator/quencher Pair8-aminonaphthalene-136-trisulfonate (ANTS)Tl+
