Measuring GPCR-Induced Intracellular Calcium Signaling Using a Quantitative High-Throughput Assay.

Shree S Kumar, Kathleen G Mountjoy

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In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

2 authors.

Shree S KumarFaculty of Medical and Health Sciences, Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Auckland, New Zealand.

Kathleen G MountjoyFaculty of Medical and Health Sciences, Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Auckland, New Zealand. kmountjoy@auckland.ac.nz.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Alterations in intracellular calcium are integral to signal transduction pathways for many G-protein-coupled receptors, but this signaling is not well studied. This is mostly due to a lack of reliable, robust, high-throughput, quantitative methods to monitor intracellular calcium concentrations in live cells. Recently, we developed a reliable, robust, quantitative method to measure intracellular calcium levels in which HEK293 cell suspensions loaded with Fura-2/AM are placed in 96-well plates. Minimum and maximum intracellular calcium levels, which are required for converting fluorescence into calcium concentrations, are calibrated using EGTA to chelate calcium and ionomycin to load calcium into cells, respectively. Fluorescence is monitored with a PHERAstar FS plate reader. We provide a detailed method for this high-throughput assay that can be used to quantitate intracellular calcium in endogenous and exogenously (stable or transient) expressed GPCRs in HEK293 cells.

Indexed as

CalciumCalcium SignalingHigh-Throughput Screening AssaysReceptors, G-Protein-CoupledFura-2HEK293 CellsHumansCalciumFura-2Receptors, G-Protein-CoupledCalcium mobilizationFluorescenceFura-2 dyeG-protein-coupled receptorsIntracellular calcium quantitationPHERAstar FS plate readerRatiometric

Identifiers

PMID39395093

DOI10.1007/978-1-62703-086-1_6

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.