Specifications

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.

Assay protocol

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

Assay details

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.

Assay validation

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.

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.