cAMP - Gi assay kits

Discover the benefits of a solution tailored for Gi coupled GPCR research

Based on HTRF technology, the kit contains all the reagents you need 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:

  • Sensitivity
  • Robustness
  • Precision
  • Ease-of-use

New to GPCR research? Cisbio has compiled a document section with all the resources you will need to get your project started, including reference documents, expert webinars, and insider tips.

Download cAMP guide - agonists of Gi/o
New Call-to-action

 

Assay Principle

Cisbio’s 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.

The protocol is simple and straightforward, with just two incubation steps:

  • Cell stimulation by the target compounds
  • cAMP detection using HTRF reagents

What to expect at the bench

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/B IC10 nM IC50 nM IC90 nM
cAMP - Gi >31 0.8 3.0 12

Product validation

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

The data below show 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 accordance with the literature.

Agonist

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 Gi cAMP Gs dynamic
Optimal Cell density / well 6000 12000
S/B 4.5 2.3
EC50 (nM) 1.6 1.9

 

Antagonist

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 Antagonists
cAMP Gi cAMP Gs dynamic
Naltrindole Naloxone Naltrindole

Naloxone

Optimal Cells density / well 6000 6000 12000 12000
S/B 4.0 4.2 2.3 2.3
IC50 (nM) 1.4 868 2.4 934

 

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
cAMP - Gi kit - 1,000 tests62AM9PEB
cAMP - Gi kit - 20,000 tests62AM9PEC
-
cAMP - Gi kit - 100,000 tests62AM9PEJ
-

Companion products

DescriptionCat. noProduct insertMSDS
cAMP - Gs Dynamic & Gi standard62AM4CDA
-
cAMP - Gs Dynamic & Gi 10 standards62AM4CDX
-
IBMX - Phosphodiesterase inhibitor - 40 µl62AMXADA
-
IBMX - Phosphodiesterase inhibitor - 500 µl62AMXADC
-
Forskolin - Adenylyl cyclase activator - 40 µl62AMYADA
Forskolin - Adenylyl cyclase activator - 800 µl62AMYADC
-
Lysis & Detection Buffer 8 - 13 ml64CL8FDD
Lysis & Detection Buffer 8 - 200 ml64CL8FDG
Stimulation Buffer 1 (5X) - 8 ml64SB1FDC
Stimulation Buffer 1 (5X) - 100 ml64SB1FDD

Literature