Gen5 version required: 1.0 or higher
Basis for the Assay:
cAMP quantification is commonly used in primary or secondary high-throughput screening for G-protein coupled receptors. Several different assay technologies can be used to quantify cAMP, including fluorescence polarization, time-resolved fluorescence (TRF), and time-resolved fluorescence resonance energy transfer (TR-FRET). This sample file provides an example of TR-FRET determination of cAMP using a LANCE™ kit from PerkinElmer® on the Synergy 2 multi-detection microplate reader.
The protocol calls for a time-resolved fluorescence measurement using a 330/80 nm excitation filter and a 665/10 nm emission filter. The time-resolved parameters are 50 μs delay and 200 μs integration time.
The experiment file contains an example of data coming from a series of standards.
11 standards are dispensed across the plate, in replicates of 16, in a 384-well plate.
The report includes a standard curve and statistics on the standard replicates.