| Assay Method Information | |
| | In Vitro Enzyme Assay |
| Description: | Human TGFβl R-1 inhibition experiments were carried out in a white 384-microplate low flange (Corning 3572) with ADP-Glo kinase Assay Kit (Promega V9101) and TGβR-1 Kinase Enzyme System (Promega V4092). Test compounds and standard Galunisertib (Cayman 15312), 50 ng/well TGFβR-1 kinase and 50 μM ATP were added in a final volumen of 10 μL/well, using Reaction buffer supplied by kit as assay buffer. The reaction mixture was incubated in gentle shaking for 120 min at RT, after incubation of 10 μL of ADP-Glo Reagent was added and incubated in gentle shaking for 40 min at RT. 20 μL of Kinase Detection Reagent was added and plate was incubated in gentle shaking for 30 min at RT. Luminiscence (1000 ms) was measured in Perkin Elmer EnSpire Multimode plate reader. |
| Affinity data for this assay | |
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