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Quantifying Gq Signaling Using the IP Homogenous Time-Resolved Fluorescence (HTRF) Assay. | LitMetric

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

Methods Mol Biol

Metabolism and Systems Science, University of Birmingham, Birmingham, UK.

Published: October 2024

AI Article Synopsis

  • * The chapter explains a specific assay, called the inositol phosphate-1 (IP) accumulation assay, which uses TR-FRET to study Gq-IP3 signaling in cells.
  • * It offers a comprehensive guide on how to set up experiments, prepare samples, and analyze data to help researchers examine the pharmacology of Gq-coupled receptors effectively.

Article 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 (IP) 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.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-4164-4_2DOI Listing

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