Assay Method Information

Assay Name:  Enzyme Inhibition Scintillation Proximity Assay
Description:  The biochemical activity of compounds was determined by incubation with wild-type or mutant p38alpha enzymes, and substrates in the presence ATP/[gamma-33P] ATP. After the reaction was stopped, the phosphorylated substrate was separated from nonincorporated radioactive ATP using SPA beads (Amersham Pharmacia Biotech), counted using TopCount (Packard) to measure substrate-incorporated phosphate. The IC50 values were calculated using a four-parameter fit of data after correction for background.
Affinity data for this assay
 

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