| Assay Method Information | |
| | PDGFRβ LanthaScreen Assay |
| Description: | Step 1. Dispensing inhibitors: Using Echo, dispense 40 nL/well (or less) compound serial dilutions in DMSO onto the assay plate.Step 2. Dispensing Kinase-Tracer solution: Add 4 μL/well Kinase-Tracer solution. Seal the plate with optically transparent plate seal. Centrifuge at 1000 rpm for 1 min.Final concentrations of components in the assay:[Tb-PDGFRβ]=0.2 nM;[Tracer 222]=40 nM;[DMSO]≤1%.Step 3. Detection: Read TR-FRET signals after 18 hours incubation at room temperature.III. Calculations and Formulas% Inhibition: % Inhibition=(NC−sample)/(NC−PC)*100 where NC is the mean of negative control (reactions without inhibitor), and PC is the mean of positive control (1 μM sunitinib).IC50 determination: Compounds are serially diluted 3-fold and tested in an 11-point dose response. IC50 values are determined from a 4-parameter fit, using the following equation: Y=Bottom+(Top−Bottom)/(1+10((Log IC50−X)*Hill slope)), where X=log10 of the compound concentration; top can be defined by PC; bottom defined by NC. |
| Affinity data for this assay | |
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