Brad Larson, B.A.1, Tracy Worzella, M.S.1, Michael Bjerke, M.S.1, Aoife Gallagher, Ph.D.2 and Eric Matthews, M.S.3
1Promega Corporation, Madison, WI, 2Deerac Fluidics, Dublin, Ireland, 3BMG Labtech
Protein kinases play a vital role in cellular activity, ranging from cell proliferation to cell death. Their substrates encompass a wide range of chemical structures such as proteins, peptides, sugars, lipids, etc. They are also implicated in many diseases, including cancer, inflammatory responses, and Alzheimer’s disease. This makes them ideal targets for high-throughput screening of potential drug targets. Because of increased demands on screening facilities, the ability to miniaturize assays has become imperative. We have developed automated Kinase-Glo® methods in low-volume 384- and 1536-well plate formats using the Deerac Fluidics Equator™ NS 808 nanoliter-scale noncontact liquid handler. Luminescent kinase activity was measured with the BMG LABTECH PHERAstar reader. The Kinase-Glo® Assay is a homogeneous method for measuring kinase activity from virtually any kinase/substrate combination by quantifying ATP remaining in solution following a kinase reaction. Excellent Z′-factor scores and accurate IC50 values in reaction volumes as low as 2μl, coupled with the extended luminescent half-life of this reagent chemistry, make this assay ideal for all high-throughput applications.