Quantifying Gq Signaling Using the IP1 Homogenous Time-Resolved Fluorescence (HTRF) Assay

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

G protein-coupled receptors (GPCRs) play pivotal roles in cellular signaling and can regulate several cellular functions such as proliferation, secretion, protein expression, and cellular metabolism. Coupling of GPCRs to members of the Gq/11 protein family results in activation of inositol trisphosphate (IP3) and accumulation of calcium intracellularly. This protocol chapter outlines a step-by-step guide for utilizing the inositol phosphate-1 (IP1) accumulation assay, a time-resolved fluorescence resonance energy transfer (TR-FRET) method, to investigate Gq-IP3 signaling. The assay serves as a valuable tool for those conducting pharmacological investigations and compound screening targeting this critical cellular pathway. This protocol chapter covers experimental setup, sample preparation, and data analysis, providing researchers with an in-depth guide to explore the pharmacology of Gq-coupled receptors.
Original languageEnglish
Title of host publicationCalcium Signaling
Subtitle of host publicationMethods and Protocols
EditorsCaroline M. Gorvin
PublisherHumana Press
Chapter2
Pages23-32
Number of pages9
Edition1
ISBN (Electronic)9781071641644
ISBN (Print)9781071641668, 9781071641637
DOIs
Publication statusPublished - 13 Oct 2024

Publication series

NameMethods in Molecular Biology
PublisherSpringer
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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