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

Karen Huss | Zendy; Pauline E. Blonigen | Zendy; Robert M. Campbell | Zendy

Open Access

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

Author(s) -

Karen Huss,

Pauline E. Blonigen,

Robert M. Campbell

Publication year - 2007

Publication title -

slas discovery

Language(s) - English

Resource type - Journals

eISSN - 2472-5560

pISSN - 2472-5552

DOI - 10.1177/1087057107300221

Subject(s) - protein kinase a , chemistry , microbiology and biotechnology , ligand binding assay , kinase , monoclonal antibody , biochemistry , antibody , biology , receptor , immunology

A Transcreener kinase fluorescence polarization (FP) assay has been developed for the serine/threonine kinase protein kinase A (PKA). The PKA Transcreener 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 PKA assay was validated with both known PKA inhibitors and library compounds. The Transcreener 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.

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