Ca2+ mobilization assays in GPCR drug discovery

Methods Mol Biol. 2015:1272:79-89. doi: 10.1007/978-1-4939-2336-6_6.

Abstract

Intracellular calcium mobilization can be measured using several methods varying in indicator dyes and devices used. In this chapter, we describe the fluorescence-based method (FLIPR Calcium 4 Assay) developed by Molecular Devices for a FlexStation and routinely used in our laboratory for detecting intracellular calcium changes. The assay is designed to study calcium mobilization induced by majority of GPCRs and calcium channels and allows for simultaneous concentration-dependent analysis of several receptor agonists and antagonists, useful in receptor characterization and drug discovery projects.

MeSH terms

  • Biological Assay
  • Bradykinin / pharmacology
  • Calcimycin / pharmacology
  • Calcium / metabolism*
  • Calcium Ionophores / pharmacology
  • Calcium Signaling
  • Drug Discovery
  • Fluorescent Dyes
  • High-Throughput Screening Assays*
  • Histamine / pharmacology
  • Humans
  • Kinetics
  • Lysophospholipids / pharmacology
  • Myocytes, Smooth Muscle / cytology
  • Myocytes, Smooth Muscle / drug effects*
  • Myocytes, Smooth Muscle / metabolism
  • Primary Cell Culture
  • Reagent Kits, Diagnostic
  • Receptors, G-Protein-Coupled / agonists
  • Receptors, G-Protein-Coupled / antagonists & inhibitors
  • Receptors, G-Protein-Coupled / metabolism*
  • Respiratory System / cytology
  • Respiratory System / drug effects
  • Respiratory System / metabolism
  • Sphingosine / analogs & derivatives
  • Sphingosine / pharmacology

Substances

  • Calcium Ionophores
  • Fluorescent Dyes
  • Lysophospholipids
  • Reagent Kits, Diagnostic
  • Receptors, G-Protein-Coupled
  • sphingosine 1-phosphate
  • Calcimycin
  • Histamine
  • Sphingosine
  • Bradykinin
  • Calcium