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Quantifying fluorescent ligand binding to GPCRs in live cells using the PHERAstar FS - a new format for HTS

Alison Carter, Kevin Thompson (1), Catherine Wark (2) 1) CellAura Technologies, 2) BMG LABTECH 06/2012

G-protein coupled receptors (GPCRs) are prominent drug targets thus demanding methods that identify GPCR ligands. CellAura Technologies developed ligands labelled with the BODIPY 630/650 fluorophore that can be used to determine IC50 values of antagonistic compounds.

 

To this end, cells are loaded with the fluorescent ligand either selective for adenosine A3 receptor or for the dopamine D1 receptor. Potential competitors are added to the cells and may displace the fluorescent ligand. Upon washing of cells, the remaining fluorescence is detected using a PHERAstar® FS in bottom read mode.

 

By means of this assay, IC50 values of adenosine and dopamine antagonists were easily determined in less than three minutes, which is a clear time-advantage compared to other fluorescent ligand binding assays taking up to 60 min.

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