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Abstract for Assay Optimization and Profiling in uHTS Format
Tracy Worzella1, Brad Larson1, Kevin Hsiao1 and Aoife Gallagher2
1Promega Corporation, 2800 Woods Hollow Road, Madison WI 53711, USA
2Deerac Fluidics, Unit 8 Enterprise Centre, Pearse Street, Dublin 2,
Ireland
We demonstrate the use of Promega 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 counterscreened 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. |
