GPCR cAMP Product Solutions

Homogeneous cAMP Detection Assay Solutions for Small Molecules and Biologics

cAMP cell-based assays measure cAMP levels in cells as a direct indication of GPCR functional status (activation or inhibition). This direct measurement is obtained by quantifying cellular cAMP accumulation levels in a dose-dependent manner with cell-based assays that are homogeneous and scalable.

Eurofins DiscoverX® provides robust, easy-to-use, and high throughput screening (HTS) HitHunter® cAMP cell-based assays that accurately detect cellular cAMP levels in a variety of applications without the need for wash steps, optimization, or specialized equipment. These versatile assays provide a simple, validated detection system, optimized with over 125 cAMP Hunter™ cell lines, and include large signal-to-background, gain-of-signal responses. They can be used to monitor GPCR activation or inhibition with cAMP Hunter, ChemiSCREEN™, or non-Eurofins DiscoverX cell lines. Easily obtain reproducible results with high sensitivity, rank order ligands based on molecular potency, or determine pharmacological profiles of your ligands.

Used in the screening of millions of data points and referenced in hundreds of peer reviewed publications ranging from basic research to clinical applications, these assays provide accurate pharmacology of your ligand or small molecule and biologics while giving you the flexibility to work with multiple cell lines.

Product Highlights

  • Optional Formats – Select from hundreds of stable cell lines, frozen ready-to-assay cells, and assay ready eXpress and bioassay kits for chemiluminescent or fluorescent assay detection
  • Assay Ready Kits – Complete, ultra-convenient kits with optimized reagents for over 125 cAMP Hunter cell lines for monitoring GPCR activation using small molecules or biologics
  • High Throughput – Simple, rapid, and scalable assays that accurately detect cellular cAMP levels in 96-well through 3456-well formats
  • Validated Applications – Referenced products in over 400 peer reviewed publications ranging from basic research to clinical applications
Consider Eurofins DiscoverX’s custom development capabilities for custom cell lines, assays, & enzyme development.

Detection Kits

  • HitHunter cAMP Assays – Accurately detect cellular cAMP levels using biologics or small molecules ligands in a variety of applications. These Enzyme Fragment Complementation (EFC) based assays monitor GPCR activation through chemiluminescent detection of cAMP production in cAMP Hunter cell lines as well as non Eurofins DiscoverX cell lines.

Assay Ready eXpress and Bioassay Kits

  • cAMP Hunter eXpress and bioassay kits – Complete assay ready kits with specific cell line, optimized reagents, and plates for quick EFC-based chemiluminescent detection. Bioassays include additional optimization for high lot-to-lot reproducibility.

Cell Lines and Ready-to-Assay Frozen Cells

  • cAMP Hunter cell lines – Naturally coupled, stable cell lines qualified to use with chemiluminescent EFC-based HitHunter® cAMP AssaysAssayComplete™ cell culture reagents, and control ligands
  • ChemiSCREEN stable cell linesparental cell lines, and ready-to-assay frozen cells – Cells based on our proprietary Chem-1 and related host cell backgrounds that contain an endogenous expression of Gα15. Gα15 is a promiscuous G protein to help funnel signaling to a common calcium readout, regardless of G protein coupling status. Use cell lines or frozen cells with chemiluminescent or fluorescent assays, and parental cell lines to generate your own GPCR stable cell lines. Additional second messenger signaling, e.g. calcium mobilization, and ERK phosphorylation, for most targets can be used to perform orthogonal assay confirmation.
  • ChemiBRITE stable cell lines and ready-to-assay frozen cells – Cells engineered to flash with added brightness upon GPCR activation and developed to provide the greatest readout flexibility, including luminescent and fluorescent calcium flux as well as cAMP. These cells express a proprietary mutant version of clytin, a calcium activated photoprotein.

Advantages of cAMP Product Solutions

Optimized for a Variety of Applications

  • Perform functional screens using small molecules or biologics (antibodies, peptides)
  • Determine pharmacological profiles and rank order ligands
  • Detect neutralizing antibodies (immunogenicity studies using serum and plasma)
  • Characterize phosphodiesterase inhibitors

HitHunter cAMP Assays

  • Eliminate optimization steps with an assay that has been designed and qualified for use with over 125 cAMP Hunter cell lines
  • Achieve precise characterization of ligand pharmacology with large assay windows, sensitive detection, and wide dynamic range for ligand bias studies and many more applications
  • Reproducible assay performance without fluorescence or serum interference and no need for specialized equipment

cAMP Hunter™ Cell Lines and Assay Ready Kits

  • Over 125 stable cell lines with unlimited culture and overexpressed naturally coupled, wild-type GPCRs
  • Ultra-convenient, complete eXpress kits for quick assessment without the need for cell culture
  • Ready-to-use bioassay kits provide cryopreserved cells and optimized reagents with high lot-to-lot reproducibility to support the lifetime of the biologic drug

ChemiSCREEN Cell Lines

  • Cell lines containing a promiscuous G protein, Gα15, to help funnel signaling to a common calcium readout, regardless of G protein coupling status
  • High functional expression of receptors with the ability to assay both agonist and antagonist in a single well
  • Stable cell lines for continuous culture and increased reliability and reproducibility assay results
  • Perform orthogonal assay confirmation such as second messenger signaling, e.g. cAMP, and ERK phosphorylation, for most targets

ChemiBRITE Cell Lines

  • Cell lines modified to flash with added brightness upon GPCR activation
  • Exceptional high signal-to-background ratio with no assay interference with autofluorescent compounds
  • Tool box capabilities allows generation of your own stable cell lines

ChemiSCREEN and ChemiBRITE Frozen Cells

  • Ready-to-Assay GPCR frozen cells come with plating media and enough cells to run either 96- or 384-well plate assays in full or half plate formats
  • Qualified for cAMP and calcium second messenger assays with results within 24 hours to accelerate your data generation

HitHunter cAMP Assay Principle

The cAMP G protein-dependent pathway involves a heterotrimeric (α/β/γ) G-protein containing a GDP molecule bound to the Gα subunit, which holds the trimer together. Upon activation, GDP is exchanged for GTP, leading to the dissociation of the Gβ/Gγ dimer from Gα. Both parts remain anchored to the membrane and become free to act upon their downstream effectors and initiate unique intracellular signaling responses. The activated Gα subunit interacts with and regulates many effector molecules such adenylyl cyclase that can ultimately lead to the accumulation of cAMP (a second messenger).

HitHunter cAMP assays are competitive immunoassays amenable for high-throughput and utilizes the Enzyme Fragment Complementation (EFC) technology where a fragment ß-galactosidase (ß-gal) enzyme donor (ED) is conjugated with cAMP. This ED-cAMP conjugate and cellular cAMP compete for binding to an anti-cAMP antibody (Ab). With low levels of cellular cAMP, most of the ED-cAMP binds to the cAMP Ab, making the ED-cAMP unable to complement with the enzyme acceptor (EA). With high levels of cellular cAMP, the anti-cAMP antibody becomes saturated allowing the ED-cAMP complex to complement with the ß-gal acceptor (EA) and form an active enzyme. The active enzyme then subsequently hydrolyzes a substrate to produce a chemiluminescent signal that is directly proportional to the amount of cAMP in the cells.

HitHunter cAMP Assays are cell-based functional, immunoassays with a chemiluminescent readout. The assays are available as complete kits that are robust, highly sensitive, and easy-to-use to study GPCR activity through cAMP production. The kits contain all the reagents needed for the detection of cAMP from whole cells expressing Gαi- and Gαs-coupled receptors induced with a biologic or small molecule ligand. The flexible assay system has been designed to work in agonist or antagonist mode for 96- and 384-well plate formats. After plating and stimulation of cells, the user simply adds the HitHunter cAMP Assay reagents to the cell following the homogeneous, simple protocol provided.

Accurately Rank Molecular Potency of Ligands

HitHunter cAMP assays provide powerful tools for drug discovery screening and revealing of the correct ligand rank order and pharmacological profile. Profiling experiment using a cAMP Hunter CHO-K1 adrenergic receptor β2 cell line to test five agonists. The data highlights the receptor’s sensitivity to the various agonists, ultimately revealing the correct rank order of the agonists and showing the agonist salbutamol (EC50 of 160 nM) is less potent than the control ligand isoproterenol (EC50 of 1.7 nM).

Easily Determine Pharmacological Profiles of Complex Assay Formats

Large assay windows and sensitive detection makes HitHunter cAMP assays ideal for studying complex assay formats like Gαi-coupled receptor antagonists. Comparative study of two antagonists of the Gαi-coupled metabotropic glutamate receptor 2 (GRM2) using a cAMP Hunter CHO-K1 GRM2 cell line. Results indicate that the Antagonist 2 (IC50 of 8 nM) is a more potent inhibitor compared to Antagonist 1 (IC50 of 24 nM).

Obtain Excellent Reproducibility and High Sensitivity for Testing Biologics

Perform quality control assays, including potency assays for lot release and stability testing in biologics. A. Evaluation of lot-to-lot consistency using a GLP agonist, GLP-1(7-36), and the cAMP Hunter Gαs-coupled GLP-1R bioassay that incorporates the HitHunter cAMP Assay for Biologics. Results indicate the assay’s excellent reproducibility for all three lots tested with S:B ratios over 10-fold and EC50’s ranging from only 76 pM to 86 pM. B. Evaluation of lot-to-lot consistency using a biologic ligand, SDF1α, and the cAMP Hunter Gαi-coupled CXCR4 receptor (chemokine C-X-C motif receptor 4) cell line. Results show high sensitivity detection and excellent reproducibility with overlapping S:B ratios of ~6 and EC50’s ranging from only 689 pM to 796 pM.

Identify Allosteric Modulators, Partial Agonists, Inverse Agonists, and Silent Agonists

HitHunter cAMP assays’ superior assay performance, with its large assay window and wide dynamic range, allows for easy identification of a diverse set of pharmacological ligands such as positive allosteric modulators (PAMs), negative allosteric modulators (NAMs), partial agonists, and more. Easily identify partial agonists. This experiment uses a cAMP Hunter Gαs-coupled glucagon receptor (GCGR) CHO-K1 cell line to analyze two agonists. Results reveal [des-His1, Glu9]- glucagon exhibits partial agonism (S:B of 9.2; EC50 of 340 nM) compared to the full native agonist, glucagon (S:B of 14.8; EC50 of 11 nM).

Application Notes Uncover Novel G-Protein or Arrestin-Biased Ligands Using a Suite of GPCR Signaling Cell-Based Assays: A Study of Biased Agonism on Opioid Receptors

As more is learned about the intricacies of GPCR signaling, the harder it becomes to accurately describe ligand activity using…

Read More
Application Notes Quantify GPCR Endocytosis and Recycling with PathHunter® GPCR Internalization Assays – Analyzing Therapeutics for Opioid and Cholecystokinin Receptors

Here we show how PathHunter GPCR Internalization assays can be used to differentiate between strongly internalizing and weakly internalizing agonist…

Read More
Application Notes Discovery of Novel G-Protein or Arrestin-Biased Ligands Using a Suite of GPCR Signaling Platforms

As more is learned about the intricacies of GPCR signaling, the harder it becomes to accurately describe ligand activity using…

Read More

Scientific Posters Accelerate Drug Development of GLP-1 & -2 Receptor Analogs with Ready-to-Use Cell-Based Assays

There is an increasing need to generate therapeutics targeting metabolic and gastrointestinal diseases to address the global burden of metabolic…

Read More
Scientific Posters Qualified, Fit-for-Purpose Bioassays for Liraglutide and Exenatide as Frozen Ready-to-Use Cells

With more than 25% of the world’s population diagnosed with type II diabetes and/or metabolic syndrome, the requirement for cost…

Read More
Scientific Posters [Wall Poster] The Druggable GPCRome

DiscoveRx offers an industry-leading portfolio of over 600 naturally coupled human and ortholog G-protein coupled receptor (GPCR) cell lines designed…

Read More
Scientific Posters Detection of Endogenous Ligand Bias: A Case Study Using the Chemokine Receptor Family

The pattern of intracellular signaling downstream of GPCR activation is now known to be specific to the ligand-receptor pair as…

Read More
Scientific Posters GLP1 NAb Assay: A Case Study for Simple, Serum-tolerant Cell-based Assays to Detect Neutralizing Antibodies to Biosimilar and Bioinnovator drugs

Neutralizing antibodies (NAbs) to biological drugs may cause loss of therapeutic effi cacy and in some cases, loss of endogenous…

Read More
Scientific Posters HitHunter® cAMP and cAMP Hunter™ Cell Line: Your Solution for Use with High Serum Samples, Difficult Gi Targets and Identification of Allosteric Modulators

The cAMP detection technologies are widely used in high throughput screening (HTS) for measuring a functional response of a GPCR…

Read More
Scientific Posters Interrogating Allosteric Interactions Using Multiple Readouts for GPCRs

Compounds that bind to allosteric sites offer many benefits over those which interact orthosterically. These include enhanced specificity, ability to…

Read More
Scientific Posters Multi-Parameter GPCR Screening: An Essential Tool for Uncovering GPCR Mutant Phenotypes

The recent appreciation of functional selectivity/ligand bias in GPCR signaling has uncovered new opportunities for therapeutic discovery and continues to…

Read More
Scientific Posters Quantifying GPCR Signaling Bias: A Simple Approach to Quantify Selectivity and Agonist Bias

Interest in GPCR ligand bias has increased in recent years due to evidence that positive and negative aspects of drug…

Read More

Target Lists GPCR Product Solutions

Your source for a complete offering of GPCR assays, cell lines, and membrane preps

Read More
Target Lists Products Target List

Comprehensive Eurofins DiscoverX product list to enable your drug discovery and development programs focused on checkpoint receptors, cytokine receptors/interleukins, cytotoxicity,…

Read More

Brochures & Flyers GPCR Reference Guide

As more is learned about the intricacies of GPCR signaling, the harder it becomes to accurately describe ligand activity using…

Read More
Brochures & Flyers Evaluate Glucose Metabolism Targeted Therapeutics

GLP Cell Lines & Ready-to-Use Assays from Characterization through QC Lot Release

Read More
Brochures & Flyers High-Throughput Screening & Lead Optimization Products

Robust Cell-Based Assays to Identify Therapeutic Hits Rapidly

Read More
Brochures & Flyers Eurofins DiscoverX – Delivering Solutions from Discovery to Market

Products & Custom Development Capabilities.

Read More
Brochures & Flyers Insights Into GPCR Drug Discovery & Development

Explore GPCR-Ligand Interactions & Signaling Pathways with Binding & Functional Assays

Read More
Brochures & Flyers Obesity & Diabetes Product Solutions

Accelerate the Development of Peptide Therapeutics for Obesity & Diabetes With MOA-Reflective Assays — Assays for Screening, Profiling, Potency, &…

Read More

Webinars Validated bioassays for metabolic & gastrointestinal diseases: Accelerating development of GLP-1 & -2 therapeutics

Therapeutics targeting glucagon-like peptide, GLP-1 and GLP-2, receptors are being developed to address globally prevalent medical disorders for managing diabetes,…

Watch Now

Videos GPCR Assays Overview

Short Overview of Eurofins DiscoverX GPCR assays. 5 min.

Watch Now
Videos GPCR Assays: Which ones to use and why for your research and drug discovery and development programs

Learn about several GPCR assays and which ones to use and why for your research and drug discovery and development…

Watch Now
Videos Custom Development Capabilities

Learn about Eurofins DiscoverX cell line engineering, assay development, and recombinant enzyme production custom capabilities. Find out more about cell…

Watch Now

Blogs & Articles Accelerating Development of GLP-1 & GLP-2 Therapeutics with Validated Bioassays

Glucagon-like peptides, GLP-1 & GLP-2, are well-researched, metabolically significant peptide hormones secreted from the L-cells of the small intestine in…

Read More
Blogs & Articles GPCR Drug Discovery & Development Insights with Dr. Terry Kenakin

GPCRs have long been regarded as a low-hanging fruit drug target as they are conveniently located on the cell surface…

Read More
Blogs & Articles Employing Assay “Volume Control” to Unveil Efficacies in Allosteric Lead Molecules for GPCRs

Drugs interact with living physiology; an in vitro assay of drug activity can be considered a snapshot of the full…

Read More
Blogs & Articles Evaluate GLP-1R Targeted Therapeutics using First-to-Market Semaglutide Qualified GLP-1R Bioassay

Glucagon-Like Peptide-1 (GLP-1), a member of the glucagon-secretin peptide family, is secreted from L-cells of the small intestine and binds…

Read More
Blogs & Articles Insights Into GPCR Drug Discovery & Development

Explore GPCR-Ligand Interactions & Signaling Pathways with Binding & Functional Assays

Read More
Blogs & Articles Four Reasons Why You Should Measure and Quantify Signaling bias in Your Molecules

Most GPCRs are pleiotropically coupled to multiple signaling pathways in cells and the availability of functional assays to separately measure…

Read More
Blogs & Articles Ligand Bias: Increasing specificity and Reducing Side Effects of Drug Compounds

Ligand bias is the ability of compounds to exert differential signal transduction responses on activation of a GPCR. This is…

Read More

White Papers The New World of GPCR Allosteric Modulation

There are at least three unique features of allosteric ligand-receptor interactions that lead to potentially valuable therapeutic behaviors…

Read More
GPCR Product Solutions

Cell-based assays to measure 2nd messengers (cAMP & calcium), β-arrestin recruitment, receptor internalization, & ligand binding

Read More
Build Your Own Assay Tools

Complete set of parental cell lines, vectors, kits, & retroparticles to build your own stable cell lines & cell-based assays

Read More
Custom Capabilities

Custom cell lines, kits, assays, & protein development capabilities optimized to fit your requirements

Read More
GPCR Calcium Product Solutions

Fluorescent assays to measure calcium mobilization in cell lines as a direct indication of GPCR or ion channel activation or…

Read More
GPCR Internalization Assays

Non-imaging, cell-based internalization assays for screening & identifying safer drugs for GPCRs

Read More
GPCR β-Arrestin Product Solutions

Universal G-protein independent cell-based assays to quantify GPCR activation based on the recruitment of β-arrestin

Read More