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Assay Optimization and Profiling in uHTS Format
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Deerac Fluidics,Promega Corp

Abstract

We demonstrate the use of Promega’s Kinase-Glo® Plus Luminescent Kinase Assay for miniaturized kinase assays using Deerac Fluidics’ Equator™ HTS liquid handling system. The Kinase-Glo® Plus Assay is a homogeneous method of measuring kinase activity by quantifying ATP in solution following a kinase reaction. The Equator™ HTS is a noncontact liquid dispenser capable of delivering volumes from 20µl down to 50nl.

Because assay development and screening are key uses for miniaturized assays, we show the Equator™ HTS can be used to optimize Kinase-Glo® reactions, and screen for kinase activity. We use the Equator™ HTS to titrate kinases and substrates to determine optimal concentrations for subsequent screening activities in 1536-well format. A panel of kinases and compounds are counter-screened for kinase selectivity profiling. 24-point serial titration is performed to determine potency. Data show that miniaturization of the Kinase- Glo® Plus Assay does not compromise assay quality. The combination of Kinase-Glo® Plus and the Equator ™ HTS provides an integrated solution for kinase drug discovery.

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