| Assay Method Information | |
| | ADP-Glo Kinase Assay |
| Description: | The STK3 kinase assay was performed in 5 μL reaction buffer containing 50 ng recombinant human STK3 protein (full length, SignalChem #S24-10G; Richmond, Canada), 250 μg/mL myelin basic protein (Sigma-Aldrich #M1891; St. Louis, MO, USA), and 50 μM ATP (Sigma-Aldrich #A7699). The STK4 kinase assay was performed in 5 μL reaction buffer containing 50 ng recombinant human STK4 protein (full length, SignalChem #S25-10G), 300 μg/mL Axltide (SignalChem #A16-58), and 50 μM ATP. IC50 values were determined with 10 concentrations of compounds serially diluted 3-fold from a starting concentration of 30 μM. Staurosporine, a non-selective protein kinase inhibitor, was included in the assay as a positive control. Three experiments were performed, each in triplicate. |
| Affinity data for this assay | |
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