Sample Files

TRF Cyclic AMP Delfia with data filters

November 29, 2012

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Related Products: Gen5 for Detection



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 TRF determination of cAMP using a Delfia® kit from PerkinElmer® on the Synergy 2 multi-detection microplate reader.

The protocol calls for a time resolved fluorescence measurement using a 340/30 nm excitation filter and a 620/40 nm emission filter, or 360/40 nm excitation filter and a 620/40 nm emission filter. The two different excitation wavelengths are provided as an example of different setups that would work. Only one excitation filter is required to run this assay. 

The experiment file contains an example of data coming from a series of standards, a control and blank wells.

Plate Configuration:

10 standards are dispensed across the plate, in replicates of 8. Column 11 contains a control in 8 replicates, and column 12 contains 8 blanks.

Report:

The report includes the standard curve and curve fitting results table, statistics on the standard replicates and the plate layout, raw data measurements and calculated concentrations in the matrix format.

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