HTRF CAMP GI KIT - 1K PTS

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

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.​

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.