| Assay Method Information | |
| | ThermoFisher SelectScreen Biochemical Kinase Profiling Service, LanthaScreen Eu Kinase Binding Assay TGFBR1 (ALK5) |
| Description: | Exemplified compounds are screened in 1% DMSO (final concentration) by 3-fold serial dilutions from 10,000 nM to 0.316 nM or at single concentrations of 10,000, 1000 and 100 nM according to the following protocol. To low volume, white 384-well plates (Greiner Cat #784207) add 160 nL 100× compound in 100% DMSO, 3.84 uL kinase buffer (50 mM HEPES, pH 7.5, 0.01% BRIJ-35, 10 mM MgCl2, 1 mM EGTA), 8 uL 2× kinase/antibody mixture in kinase buffer (final ALK5 concentration of 5 nM; final Eu-anti-GST antibody concentration of 2 nM), 4 uL 4× Tracer #178 in kinase buffer (final concentration of 5 nM). Gently shake the plates for 30 seconds and incubate at room temperature for 1 hour before reading fluorescence on a plate reader. Percent inhibition is determined compared to DMSO only control (maximal ALK5 tracer binding) and sigmoidal dose response curve fit yielded IC50 values as indicated in Table 2. |
| Affinity data for this assay | |
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