| Assay Method Information | |
| | Inhibition Assay |
| Description: | Using human p38 MAPK alpha , the p38 MAPK inhibitory activity of Compound (I) was studied by a method partially modified from the method described in Current Medicinal Chemistry (2004, No. 11, pp. 721-730).A solution of each test compound solution in 100% DMSO and a p38alpha /SAPK2a solution (Final Concentration: 1.5 nM) (Invitrogen) were added to a 384-well plate, and then the plate was incubated at room temperature in a dark place for 1 hour. Thereafter, ATP (Final Concentration: 100 uM), which is a phosphate donor, and biotinylated ATF2 (Final Concentration: 30 nM) (upstate), which is a substrate, were added, and the resulting mixture was allowed to react at room temperature in a dark place for 1 hour (the final concentration of DMSO was 0.25%). After the reaction, an anti-phosphorylated ATF2 antibody (Final Concentration: 1 nM) (Cell Signaling), anti-IgG acceptor beads (Final Concentration: 20 ug/mL) (PerkinElmer) and streptavidin donor beads. |
| Affinity data for this assay | |
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