A Functional Assay for GPR55: Envision Protocol

Sharon Anavi-Goffer; Ruth A Ross

doi:10.1007/978-1-4939-3539-0_8

A Functional Assay for GPR55: Envision Protocol

Methods Mol Biol. 2016:1412:77-83. doi: 10.1007/978-1-4939-3539-0_8.

Authors

Sharon Anavi-Goffer^{1

2}, Ruth A Ross^{3

4}

Affiliations

¹ Department of Behavioral Sciences, Ariel University, Ariel, 40700, Israel. s.goffer@ariel.ac.il.
² Institute of Medical Sciences, University of Aberdeen, Aberdeen, UK. s.goffer@ariel.ac.il.
³ Institute of Medical Sciences, University of Aberdeen, Aberdeen, UK.
⁴ Department of Pharmacology and Toxicology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada.

PMID: 27245893
DOI: 10.1007/978-1-4939-3539-0_8

Abstract

AlphaScreen(®) SureFire(®) assay is a novel technology that combines luminescent oxygen channeling technology, nano-beads, and monocloncal antibodies to detect the level of a selected protein in a volume lower than 5 μl. This method is more sensitive compared with the traditional enzyme-linked immunosorbent assays (ELISA), and can detect an increasing number of new targets. Here, we described a method for AlphaScreen(®) SureFire(®) assay that targets ERK1/2 phosphorylation, a primary downstream signaling pathway that conveys activation of GPR55 by L-α-lysophosphatidylinositol (LPI) and certain cannabinoids.

Keywords: AlphaLISA; AlphaScreen; Cannabinoid receptor; Enspire; GPR55; High-throughput screening; Phytocannabinoids.

MeSH terms

Biological Assay*
Cell Culture Techniques
HEK293 Cells
Humans
MAP Kinase Signaling System
Phosphorylation
Receptors, Cannabinoid
Receptors, G-Protein-Coupled / metabolism*

Substances

GPR55 protein, human
Receptors, Cannabinoid
Receptors, G-Protein-Coupled

Abstract

MeSH terms

Substances

Grants and funding