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Journal of Biomolecular Screening
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Determination of Equilibrium Dissociation Constants in Fluorescence Polarization

Linda Prystay

PerkinElmer Life Sciences, Montreal, Quebec, Canada

Mylene Gosselin

PerkinElmer Life Sciences, Montreal, Quebec, Canada

Peter Banks

PerkinElmer Life Sciences, Montreal, Quebec, Canada

A simple mathematical model (the FP Kd model) is used to generate the dissociation equilibrium constant (Kd) for G protein-coupled receptor-ligand binding measured using fluorescence polarization (FP) saturation curve analysis. The model generates data that may be analyzed by the method of Scatchard. The validity of the FP Kd model is proven in six model systems in which the modeled Kd values are within a factor of 5 of inhibitory equilibrium constant values obtained from radioligand competition assays.

Journal of Biomolecular Screening, Vol. 6, No. 3, 141-150 (2001)
DOI: 10.1177/108705710100600304
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