Assay Method Information |
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| Enzyme Compound Activity (IC50) and Cell Compound Activity (IC50/EC50) |
Description: | A fluorescence polarization assay was used to assess the ALK5 binding capacity and biochemical activity of compounds. ALK5 protein was added to each well of a 384 well assay plate containing test compounds, FP probe, and ATP. Following incubation for 30 min at room temperature, fluorescence polarization was measured on a BMG Labtech PHERAstar plate reader using a 650/690 nm optic module. mP values were then used to calculate IC50 values for each compound tested. IC50/EC50 values were obtained from measurements of compound cellular activity to inhibit nuclear translocation of a GFP-2/Smad2 fusion reporter in MDA-MB-468 cells. |
Affinity data for this assay | |
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