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Member since: 2011
Organization: Center, Computational Sci
"we analyze data generated from this assay."
Universal, HTS Kinase Screening Assays
How It Works
The kinase reaction is conducted under the appropriate conditions. ATP remaining at the time that the Kinase-Glo® Reagent is added is used as a substrate by the Ultra-Glo™ Luciferase to catalyze the mono-oxygenation of luciferin and the generation of light. Luminescence is inversely related to kinase activity.
Linear out to 500μM ATP
The Kinase-Glo® Platform consists of three assay formats: the Kinase-Glo® Assay, which is used to monitor kinase activity using up to 10μM ATP; the Kinase-Glo® Plus Assay, which is used for assays requiring higher ATP concentrations (up to 100μM); and the Kinase-Glo® Max Assay, which is used for assays requiring up to 500μM ATP.
Perfect for HTS applications
Z´-factor is a statistical value that compares the assay dynamic range to data variation in order to assess assay quality. Z´-factors greater than 0.5 indicate excellent assay quality.
Uncover non-ATP binding site inhibitors
Use the Kinase-Glo Assays to distinguish ATP competitive inhibitors from noncompetitive inhibitors. Use PKA inhibitor (noncompetitive, PKI, and competitive, H89) titrations were performed using the Kinase-Glo Plus Assay. IC50 results determined using Kinase-Glo Plus varied just 2-fold for the noncompetitive inhibitor PKI (3.5nM vs. 7.9nM) at 10 and 100μM ATP, respectively. The IC50 results varied 7-fold for the competitive inhibitor H89 (0.06μM vs. 0.4μM) at 10 and 100μM ATP, respectively.