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Development of a Transcreener^TM Kinase Assay for Protein Kinase A and Demonstration of Concordance of Data with a Filter-Binding Assay Format

Lilly Research Laboratories A Division of Eli Lilly and Company, Indianapolis, Indiana

Pauline E. Blonigen

Lilly Research Laboratories A Division of Eli Lilly and Company, Indianapolis, Indiana

Robert M. Campbell

Lilly Research Laboratories A Division of Eli Lilly and Company, Indianapolis, Indiana

A Transcreener^TM kinase fluorescence polarization (FP) assay has been developed for the serine/threonine kinase protein kinase A (PKA). The PKA Transcreener^TM kinase assay is an homogenous, competitive antibody-based FP assay that uses Far Red Alexa Fluor 633-labeled adenosine 5' disphosphate (ADP) tracer and mouse monoclonal anti-ADP antibody. The Transcreener^TM PKA assay was validated with both known PKA inhibitors and library compounds. The Transcreener^TM PKA assay is resistant to low-wavelength (or common) fluorescent interference from small-molecule library compounds and generates IC₅₀ results comparable with current radioactive filter-binding assay. (Journal of Biomolecular Screening 2007:578-584)

Key Words: Transcreener^TM • protein kinase A • fluorescence polarization • Alexa Fluor 633

This version was published on June 1, 2007

Journal of Biomolecular Screening, Vol. 12, No. 4, 578-584 (2007)
DOI: 10.1177/1087057107300221


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