Miniaturized GloSensor™ cAMP Assay Using the Echo® Liquid Handler Poster
Part # PS153
Tracy Worzella1, Tim Allison2, Pete Stecha1, Kevin Kershner1, Mei Cong1, Chad Zimprich1 and Brock Binkowsk1
1Promega Corporation, Madison, WI USA; 2, Labcyte Inc. Sunnyvale, CA USA
As biological assays used for HTS and uHTS are miniaturized, it is critical to maintain assay sensitivity while minimizing reagent volumes and maximizing throughput. The GloSensor™ cAMP Assay provides an extremely sensitive and easy to use, real-time luminescent assay format for the interrogation of overexpressed or endogenous GPCRs that signal via changes in the intracellular concentration of cAMP. Tipless, touchless transfers with the Echo® liquid handler eliminate the need for costly disposable tips and greatly simplify assay development efforts. Precise and accurate drop placement eliminates cross-contamination. In this work, we demonstrate optimization of the GloSensor™ cAMP Assay in low-volume format using a stably transfected HEK293 cell line model expressing the melanocortin 4 receptor (MC4R) and a cAMP-sensing variant of firefly luciferase. We show that the Echo® liquid handler is able to titrate small molecule and peptide agonists and antagonists, providing robust assay results that are achieved with significantly reduced volumes of cells and compound.
Printed in USA.