Reactions are rapid, taking as little as two minutes to reach equilibrium, whereupon binding can be detected immediately. The application of fluorescence polarization technology to homogeneous receptor binding assays is based on a decrease in the speed of molecular rotation of a ligand or fluo-peptide that occurs upon binding to a larger molecule, such as a receptor protein, allowing for polarized fluorescent emission by the bound peptide. This concept has been proven, well validated and can be applied to HTS receptor binding assays. Fluorescence polarization is empirically determined by measuring the vertical and horizontal components of fluorophore emission following excitation with plane polarized light.
Polarized emission is increased when the molecular rotation of a fluorophore is reduced.
The magnitude of the polarized signal relates quantitatively to the extent of fluorescent ligand binding. FP assay yields equivalent MC3 agonist rank order potencies as published MC3 data using radioligand as tracer. 2 Test Kits cover a range of characterized receptor-ligand combinations developed and validated in fluorescence polarization based binding assays. Ideal for researchers who want to test this platform in their own lab and to gain experience implementing and optimizing a FP based binding assay. Special pricing is possible based on the number of compounds screened.