| Assay Method Information | |
| | PDGFRα Kinase Assay |
| Description: | The PDGFRα enzyme assays were conducted per the manufacturer's directions (Promega PDGFRα Kinase Enzyme System and ADP Glo Assay Cat #V8011). Briefly, exemplary PDGFRα inhibitor compounds described herein were assayed in an 11-point dose response curve with a maximum concentration of 10,000 nM and 3-fold dilutions to a minimum concentration of 0.169 nM. 20 ng of purified PDGFRα protein was added to the test article. Subsequently, 150 uM of ATP and 1 μg of substrate, Poly (Glu4Tyr1) was added to each reaction, and the PDGFRα kinase mediated conversion of ATP (adenosine triphosphate) to ADP (adenosine diphosphate) was allowed to continue for two hours. ADP-Glo was added to halt the kinase reaction and deplete the remaining ATP. Kinase detection reagent, containing luciferase and luciferin, was used to convert the ADP signal into luminescence. Luminescence was measured using a Molecular Devices Spectramax ID5 and compared to a standard curve to determine kinase activity. IC50s were calculated using Prism 9, Graphpad software. |
| Affinity data for this assay | |
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