| Assay Method Information | |
| | P38alpha Kinase Inhibition Assay |
| Description: | Enzymatic activity assay was performed in 96-well microtiter plates. Various concentrations of the test compound or vehicle controls were preincubated for one hour with the human p38alpha (SAPKa) enzyme. The reaction started by addition of biotinylated ATF2 substrate and ATP. After incubation at 25 deg C for 1 hr. The detection reagents, streptavidin-XL665 and antiphosphoresidue antibody coupled to Europium cryptate, caused the juxtaposition of the cryptate and the XL665 fluorophore, resulting in fluorescence energy transfer (FRET). The FRET intensity depends on the amount of bounded cryptate antibody, which is proportional to the extent of substrate phosphorylation. FRET intensity was measured using Victor 2 V spectrofluorometer. Data were analyzed by nonlinear regression (Hill equation) to generate a dose-response curve. The calculated IC50 value is the concentration of the test compound, which caused a 50% decrease in the maximal FRET intensity. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |