Abstract
Activation of G(q) protein-coupled receptors can be monitored by measuring the increase in intracellular calcium with fluorescent dyes. Recent advances in fluorescent kinetic plate readers and liquid-handling technology have made it possible to follow these transient changes in intracellular calcium in a 1,536-well plate format for high-throughput screening (HTS). Here, we have applied the latest generation of fluorescence kinetic plate readers to multiplex the agonist and antagonist screens of a G protein-coupled receptor (GPCR). This multiplexed assay format provides an efficient and cost-effective method for HTS of G(q)-coupled GPCR targets.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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CHO Cells
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Calcium / analysis
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Calcium / metabolism*
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Calibration
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Cricetinae
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Cricetulus
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Cyclic AMP / metabolism
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Dose-Response Relationship, Drug
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Drug Evaluation, Preclinical
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Fluorescent Dyes
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Indicators and Reagents
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Kinetics
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Muscarinic Agonists / pharmacology
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Muscarinic Antagonists / pharmacology
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Radioligand Assay
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Receptor, Muscarinic M1 / drug effects
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Receptors, G-Protein-Coupled / agonists*
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Receptors, G-Protein-Coupled / antagonists & inhibitors*
Substances
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Fluorescent Dyes
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Indicators and Reagents
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Muscarinic Agonists
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Muscarinic Antagonists
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NPSR1 protein, human
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Receptor, Muscarinic M1
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Receptors, G-Protein-Coupled
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Cyclic AMP
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Calcium