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| | GSK-3beta Kinase Inhibition Assay |
| Description: | Human GSK-3beta was expressed as the N-terminal FLAG-tagged protein using a baculovirus expression system. The kinase assay was was conducted in a 96-well plate assay format. The kinase reactions were initiated by addition of ATP-solution (final 500 nM), and were incubated for 90 min at 37 deg C for GSK-3 beta. The reactions were terminated by the addition of Kinase-Glo reagent containing EDTA. Ten minutes after addition of Kinase-Glo reagent, the luminescence was measured on a Wallac ARVO 1420 instrument (PerkinElmer, Shelton, CT). The reaction window was calculated from the difference of the average signals obtained from the control (5% DMSO) and the background wells. The inhibitory activity of compounds is expressed by the inhibitor concentration that produced 50% inhibition (IC50) of the enzyme activity in the absence of inhibitor. The IC50 values were obtained by linear regression analysis with a GraphPad Prism (version 3.02 for Windows, GraphPad software, Inc. San Diego, CA). The best-fit lines were obtained by analyzing the logistic fitting equation. |
| Affinity data for this assay | |
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