Technology > GPCRs > Arrestin

GPCR Technology – β-Arrestin

PathHunter® β-Arrestin GPCR assays are whole cell, functional assays that directly measure GPCR activity by detecting the interaction of β-Arrestin with the activated GPCR. Because Arrestin recruitment is independent of G-protein signaling, these assays offer a powerful and universal screening and profiling platform that can be used for virtually any Gi-, Gs, or Gq-coupled receptor.
Pathhunter GPCR Arrestin
 

Assay Principle

In this system, the GPCR is fused in frame with the small, 42 amino acid fragment of β-gal called ProLink™ and co-expressed in cells stably expressing a fusion protein of β-Arrestin and the larger, N-terminal deletion mutant of β-gal (called enzyme acceptor or EA). Activation of the GPCR stimulates binding of β-Arrestin to the ProLink-tagged GPCR and forces complementation of the two enzyme fragments, resulting in the formation of an active β-gal enzyme. This action leads to an increase in enzyme activity that can be measured using chemiluminescent PathHunter® Detection Reagents.

The PathHunter® Arrestin Advantage

  • Validated with over 180 GPCRs – compatible with all GPCR classes (A, B and C), regardless of interacting G-protein (Gs, Gi/o and Gq/11)
  • Single readout - independent of G-protein coupling, eliminates need for multiple assays
  • Universal format – independent of G-proteins, ideal for orphan GPCR screening
  • Stable cell lines - increased reliability and reproducibility
  • Stoichiometric signal generation - accurate compound efficacy measurements
  • Standardized assay conditions - efficient comparison studies across receptors or receptor families
  • Specific signal – tagged receptor eliminates background from endogenous receptors
  • Screening-friendly format – superior assay windows and robust performance


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