Assay Method Information

Assay Name:  In Vitro Assay for Detecting and Measuring Modulation of hFPR1 by Compounds
Description:  Cyclic Adenosine Monophosphate (cAMP) Assay: CHO-K1 cells stably overexpressing hFPR1 were plated into 384-well cell culture plates (2,000/well/5 μL) in 1× Stimulation Buffer (LANCE Ultra cAMP Kit, Perkin Elmer, TRF0263). 1 μL compound solution (prepared in ddH2O containing 0.2% DMSO) was added to each well. Plates were centrifuged briefly, and cells were incubated at 37° C. in 5% CO2 for 10 minutes. 4 μL solution containing 2.5 μM Forskolin and 0.25 nM WKYMVm (an FPR1 agonist, GLPBIO, GC15140) was then added to each well. 5 μL of Eu-cAMP solution (diluted to manufacturer's recommended working concentration in Detection Buffer, LANCE Ultra cAMP Kit) and 5 μL of ULight-anti-cAMP solution (diluted in Detection Buffer, LANCE Ultra cAMP Kit) were added to each well. Plates were centrifuged briefly, and cells were incubated at room temperature for 1 hour. Fluorescence emission at 665 nm and 620 nm from the samples was measured by a Perkin Elmer Envision instrument.
Affinity data for this assay
 

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