cAMP Gi kit detects changes of cAMP accumulation in response to Gi coupled GPCR activation or inhibition
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  • Ease-of-use
  • High sensitivity
  • Accurate pharmacology
cAMP Gi kit detects changes of cAMP accumulation in response to Gi coupled GPCR activation or inhibition
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In stock

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 Gi-activation, leveraging an optimized combination of robustness and sensitivity.

Assay principle

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

Assay protocol

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

Product specifications

Designed with signal performance in mind, the kit offers a tailored standard curve ideally suited to the study of Gi coupled GPCRs.

S/BIC10 nMIC50 nMIC90 nM
cAMP - Gi>310.83.012

Introduction to product validation

The cAMP Gi kit allows enhanced performances for Gi coupled receptor assessment.

The data below shows the pharmacological validation of the Gi kit for the stably expressing delta opioid receptor cell line. Agonists and antagonists were well characterized, showing potency in agreement with the literature.

Validation on agonist dose response

​CHO cells stably expressing the delta opioid receptor (DOR) at 6000 cells/well were treated with the SNC-162, a reference agonist, for 45 min.

SNC-162 was well characterized, displaying the right potency in good correlation with the literature.​ 

                                    

    
DOR Agonist SNC-162
   
cAMP GicAMP Gs dynamic
Optimal Cell density / well600012000
S/B4.52.3
EC50 (nM)1.61.9

Validation on antagonist dose response

CHO cells stably expressing the delta opioid receptor (DOR) at 6000 cells/well were treated with the SNC-162 agonist for 45 min at the EC90 value (10 nM), which is the optimal concentration to use for the antagonist mode experiment.

Fitting the serial dilutions of the Naltrindole and the Naloxone antagonists enabled IC50 calculation of the compounds (potency), which was in perfect correlation with the literature.


DOR AntagonistscAMP GicAMP Gs dynamic
NaltrindoleNaloxoneNaltrindole

Naloxone

Optimal Cells density / well600060001200012000
S/B4.04.22.32.3
IC50 (nM)1.48682.4934


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