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Assay Method Information

Assay Name:  Luminescence Cell-Free Homogeneous Dose Retest to Identify Inhibitors of Glycogen Synthase Kinase-3 beta Activity
Description:  Keywords: GSK3beta, dose response, kinase, inhibition, HTS Assay Overview: The glycogen synthase kinase-3 beta (GSK-3b) is a known master regulator for several cellular pathways and plays a critical role in metabolism, transcription, development, cell survival, and neuronal functions. The overall objective is to identify one or multiple series of inhibitors of GSK-3beta with micromolar potency. We had conducted a primary screen and now this assay is to determine the potency of the screen activities by testing these actives in doses. Basically, 1.6 ng of GSK3beta (as a GST fusion from BPS Bioscience) was incubated with compounds in doses in the presence of 25 uM of ATP (specific reaction conditions see Protocol) for 60 minutes at ambient temperature in 1536 plates (Aurora 29847). The kinase activity was measured with ADP-Glo (Promega V9103) and signals were read with Viewlux (PerkinElmer). Positive control (GW8510 at 20 uM) was included in each plate and used to scale the data in
Affinity data for this assay
 

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Last update November 1, 2007
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