Thursday, August 16, 2012

Inside Applications: Quantifying Fluorescent Ligand Binding to GPCRs in Live Cells using the PHERAstar FS - a new format for HTS

Fluorescent ligand binding
Fluorescent ligand binding at human dopamine D1 receptors, and displacement with the unlabelled D1-selective competitor SCH23390.
Point Fluorescent ligand binding to GPCRs in live cells measured using a PHERAstar FS
Point Unlabelled competitor displacement IC50 curves quantified in 96-well format in
< 3 minutes per plate
Point A fast alternative to High Content Analysis for receptor binding and screening
Fluorescent GPCR dopamine and adenosine ligands (developed by CellAura Technologies Ltd) were quantified with the help of the PHERAstar FS from BMG LABTECH. The adjustable focal height feature and the advanced bottom reading capability of this microplate reader were utilized to measure the adherent cell layer at the bottom of the well. Real-time 96-well fluorescence measurements were performed "on the fly" and resulted in read times < 3 minutes per plate. This is in stark contrast to High Content Screening (HCS) assays, which can take around 60 minutes to achieve the same endpoint.

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