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cAMP Gs dynamic kit HTRF®

This HTRF kit detects changes of cAMP accumulation in response to Gs coupled GPCR activation or inhibition

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  • Ease-of-use Ease-of-use
  • High sensitivity High sensitivity
  • Accurate pharmacology Accurate pharmacology

This HTRF kit detects changes of cAMP accumulation in response to Gs coupled GPCR activation or inhibition

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Overview

Based on HTRF technology, the kit contains all the reagents needed to quantify cAMP, ensuring that you benefit from a complete tool for your needs.

Cisbio's cAMP kits are best suited for measuring cAMP modulations in response to Gs-activation, leveraging an optimized combination of robustness and sensitivity

Benefits

  • SCREENING OF GS COUPLED GPCR
  • LEAD OPTIMIZATION OF GS COMPOUNDS
  • STUDY OF GS GPCR RESPONSES

Assay principle

cAMP kits are based on a competitive format involving a specific antibody labeled with cryptate (donor) and cAMP coupled to d2 (acceptor). This enables the direct characterization of all types of compounds acting on Gs-coupled receptors in either adherent or suspension cells. Native cAMP produced by cells competes with d2-labeled cAMP for binding to monoclonal anti-cAMP Eu3+ cryptate.
Diagram of cAMP Gs Dynamic assay principle

Assay protocol

The protocol is simple and straightforward, with just two incubation steps: - Cell stimulation by the target compounds - cAMP detection using HTRF reagents
Diagram of cAMP Gs Dynamic assay protocol

cAMP Gs dynamic product specifications

Use this optimized kit to study your most challenging and precious cellular models.

Designed to produce results with minimal cell consumption, this kit features the lowest nanomolar IC50.

Left-shifted IC50 enables you to quantify lower cAMP concentrations, significantly reducing the number of cells required.

cAMP Gs dynamic standard curve characteristics

S/BIC20 nMIC50 nMIC80 nM
cAMP - Gs dynamic>140.914.0718.1
cAMP - Gs HiRange>284.2722.8121

Validation on agonist dose response

HEK293 cells expressing the endogenous ß2 adrenergic receptor were treated with the isoproterenol agonist for 45min.

Isoproterenol was well characterized, displaying the right potency in good correlation with the literature.

Isoproterenol agonist does response on the B2 adrenergic receptor

Validation on antagonist dose response

HEK293 cells at 7,500 cells/well (optimal cellular concentration situated in the dynamic range of the kit) were treated with a serial concentration of isoproterenol, a beta 2 adrenergic receptor full-agonist.

The EC80-EC90 was determined to choose the optimal agonist concentration to use for the antagonist mode experiment.

Fitting the serial dilution of the antagonist allowed IC50 calculation for the compounds (potency) which was in perfect correlation with the literature.

Propanolol antagonist does response on the B2 adrenergic receptor

Structure, function and pharmacology of G protein-coupled receptors: State-of-the-art and future challenges

In collaboration with the Faculty of health and medical sciences Denmark - Scientific Presentations

HTRF assays of IP1, cAMP and pERK pathways employed to study biased signaling of the calcium-sensing receptor

In collaboration with the Faculty of health and medical sciences Denmark - Scientific Presentations

HTRF a versatil approach for 7TM drug discovery

In collaboration with MRC - Scientific Presentations

cAMP Fresh and Frozen Cell Usage

In collaboration with Bristol Myers Squibb - Scientific Presentations

IP-One & cAMP HTRF Functional Cell-based Screening

In collaboration with Schering AG - Scientific Presentations

Biomarker and Cell Signaling Assays for Fibrosis and NASH

HTRF and Alpha solutions for NASH - Flyers

GPCR research from A to Z

HTRF products and supporting scientific content to investifgate GPCRs - Flyers

Application of Bioassay Performance Monitoring Process with Statistical Tools

In collaboration with Amylin Pharmaceuticals Inc., Biocon Limited - Posters

Characterisation of Inositol Phosphate & cAMP Signalling from GPCRs using HTRF

In collaboration with College of Science, King Saud University, University of Western Australia - Posters

HTRF® technology on the SpectraMax® i3 Multi-Mode Platform

In collaboration with Mollecular Devices - Posters

Differentiation of partial and full GPR40 agonists for the treatment of type 2diabetes

In collaboration with Jansen research & development - Posters

Ultimate guides to master Gs and Gi GPCR assays

Optimizing cAMP assays has never been easier - Guides

Understanding GPCRs is the key to improved DD

In collaboration with GEN - Guides

How to run a cAMP HTRF assay

What to expect at the bench - Videos

Product Insert cAMP Gs dyn / 62AM4PEB-62AM4PEC-62AM4PEJ

62AM4PEB-62AM4PEC-62AM4PEJ - Product Insert

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

GPCR in Cardiovascular Research: The Relevance of Camp and IP-1 Second Messenger Assays

See how peer researchers monitor GPCR signaling with HTRF - Application Notes

Molecular basis of neuroinflammation and neurodegeneration diseases

The essential guide for extending your knowledge on the molecular mechanisms of neurodegenerative diseases - Guides

Neurodegeneration and its main related diseases

Discover this infographic design on neurodegenerative diseases - Infographics

GPCRs Pharmacology

A comprehensive overview of pharmacology's main ligands - Guides

Three drug discovery approaches for Multiple Sclerosis

Power up your multiple sclerosis research projects with this technical note - Application Notes

GPCR compounds identification and pharmacological characterization with GTP Gi binding assay

Characterization of candidate molecules through GTP binding assays - Application Notes

Upstream/Downstream GPCR Readout Monitoring with GTP Gi Binding and cAMP Gi HTRF Assays

Read this note featuring two case studies that demonstrate the relevance of combining upstream (GTP) and downstream (cAMP) readouts for characterizing pharmacological compounds - Application Notes

GPCRs: the pathway to Discovery

Complete solutions for GPCR Drug Discovery - Brochures

Cyclic-AMP: A Functional Second Messenger for Assaying GPCR Target Activation

Selection of kits to detect of a broad range of cAMP - Application Notes

Safety Data Sheet cAMP Gs dyn / 62AM4PEB

62AM4PEB - Safety Data Sheet

Safety Data Sheet cAMP Gs dyn / 62AM4PEC

62AM4PEC - Safety Data Sheet

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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