PathHunter™ eXpress β-Arrestin GPCR Kits

Universal GPCR Technology for Direct Pharmacology in Live Cells

The PathHunter™ eXpress β-Arrestin GPCR assays are based on the proven PathHunter™ β-Arrestin technology which is a universal method for detection of GPCR activity regardless of coupling status. Unlike second messenger or other imaging assays, the PathHunter β-Arrestin assay is unique in that it provides a direct measure of β-Arrestin binding to the GPCR of interest and can be used with any Gi-, Gq-, Gs-coupled receptor.

Product Features

•Over 130 human and mouse Gi-, Gq-, and Gs-coupled GPCRs available
•Off-the-shelf complete kit format
•Pre-validated cells and reagents
•Complete step-by-step protocol
•Assay-Ready frozen cells
•Read on any standard benchtop luminometer

How the PathHunter™ eXpress β-Arrestin GPCR Assay Works

In this system, β-Arrestin is fused to an N-terminal deletion mutant of β-gal (termed the enzyme acceptor, EA) and the GPCR of interest is fused to a smaller (42 amino acids), weakly complementing fragment termed ProLink™. In cells that stably express these fusion proteins, the interaction of β-Arrestin and the GPCR following ligand stimulation forces the complementation of the two β-gal fragments resulting in the formation of a functional enzyme that converts substrate to detectable signal. In the absence of an interaction between the GPCR and β-Arrestin, the enzyme activity is low due to the low affinity of the two enzyme fragments.

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Figure 1. Confirm pharmacology of your compound regardless of whether it is an agonist, antagonist, inverse agonist or allosteric modulator using a simple, homogeneous protocol combined with chemiluminescent detection.

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Figure 2. Performance comparison between live cell and frozen eXpress cells
(A) Correlation between EC₅₀ values of control agonists obtained using live cells and PathHunter eXpress cells. Data is presented for 100 receptors. EC₅₀ values above the line indicate higher sensitivity in the eXpress format compared to live cells. (B) CHO-K1 PathHunter eXpress cells
and live cells expressing the Somatostatin 2 (SSTR2) receptor were stimulated with increasing concentrations of SST-28, a known agonist, for 90 minutes prior to the addition of detection reagents. PathHunter eXpress cells show equivalent pharmacology to live cells.

Kit Contents

•Assay-ready, frozen PathHunter™ eXpress cells (2 vials each, 1 x 10⁶ cells/vial)
•Optimized Assay Buffer
•PathHunter™ Detection Reagents
•96-well Tissue Culture Treated Plates

Available Collaterals

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