Enzyme Inhibition Assay: The kinase enzyme binding activities of compounds disclosed herein were determined using a proprietary assay which measures a...

ID: ALA3706164

Type: Binding

Description: Enzyme Inhibition Assay: The kinase enzyme binding activities of compounds disclosed herein were determined using a proprietary assay which measures active site-directed competition binding to an immobilized ligand (Fabian, M. A. et al., Nature Biotechnol., 2005, 23:329-336). These assays were conducted by DiscoverX (formerly Ambit; San Diego, Calif.). The Kd value (Dissociation constant value) was calculated as the index of affinity of the compounds to each kinase. The enzyme inhibitory activities of compounds disclosed herein were determined by FRET using synthetic peptides labelled with both donor and acceptor fluorophores (Z-LYTE, Invitrogen Ltd., Paisley, UK). The inhibitory activities of test compounds against the p38 MAPKalpha isoform (MAPK14: Invitrogen), were evaluated indirectly by determining the level of activation/phosphorylation of the down-stream molecule, MAPKAP-K2. The p38 MAPKalpha protein (80 ng/mL, 2.5 uL) was mixed with the test compound (2.5 uL of either 4 ug/mL, 0.4 & 0.04 ug/mL).

Format: BAO_0000357

Organism: Homo sapiens

Target:  MAP kinase p38 alpha(ALA260)

Document:  ALA3639240

BindingDB Source:  6955