Cisbio HTRF assay reading parameters, dose-response curves and Z’-factor calculation

Related Application Notes: Automated, High Throughput Methods for the Quantification of Endogenous Cellular Kinase Activity using HTRF Technology and Automated, High Throughput, HTRF®-Based Detection of Histone Methyltransferase and Demethylase Enzyme Activity

Gen5 v2.01 required

HTRF technology can be used to quantify the activity and modulation of a number of important drug targets. These include, but are not limited to, GPCRs, kinases, epigenetic, and biological therapeutic proteins. The time-resolved fluorescence resonance energy transfer (TR-FRET) readout allows for the generation of accurate experimental data by eliminating the emission signal from most auto-fluorescent compounds, and the incorporation of a ratiometric readout. The HTRF detection module typically incorporates either a Eu3+- or Tb-cryptate donor molecule. This sample file illustrates the typical reading parameters for HTRF when using a Eu3+-cryptate donor molecule, along with dose-response curves and Z’-factor calculation.