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Assay Method Information

Assay Name:  JAK3 ELISA-based Kinase Activity Assay
Description:  In this ELISA-based kinase activity assay, a 96-well assay plate is coated with an artificial polypeptide serving as kinase substrate that contains tyrosineresidues being phosphorylated by JAK3 in presence of ATP. For the kinase reaction, the recombinant active human JAK3 protein fragment (aa 781 to 1124) is incubated with different concentrations of the inhibitor candidate and an appropriate amount of ATP (twice the Km value). As a consequence, the polypeptidic kinase substrate is phosphorylated as a function of inhibitory potency of the test compound. Phosphorylated tyrosine residues are detected via amonoclonal phopho-tyrosine HRP- conjugated antibody and quantification of phosphorylation degree is perfomed by comparing the phosphorylation achieved in absence (positive control) and presence of inhibitor candidate.
Affinity data for this assay
 

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Last update November 1, 2007
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