Cell based functional assays


cAMP (Glo)
Adenosine 3`, 5` cyclic monophosphate (cAMP) is a omnipresent  cellular second messenger that plays  a key role in signal transduction process linking membrane G-protein coupled receptors (GPCRs) to their ligands thus activating intracellular enzymatic activity and gene expression. cAMP is synthesized from ATP by membrane-bound adenylate cyclase. Binding of ligands or hormones to their specific GPCRs activates GTPs binding proteins (Gs or Gi) which either stimulate or inhibit adenylate cyclase. Concentration of cAMP inside the cell is a function of the ratio of the rate of its synthesis from ATP by adenylate cyclase and breakdown to AMP by specific phosphodiesterases.

In the light of the fact that GPCRs constitute one of the largest gene families of signaling proteins research connected with those membrane proteins appears to be of great significance especially in the area of central nervous system (CNS).

Biocentrum has an extensive experience in analysis of GPCRs activation upon binding of extracellular ligands and offers his knowledge in service of characterization of GPCRs stimulation.


 Image

(A) Agonist dose response curve. CHO cells with stable expression of dopamine D2 receptor were incubated with forskolin and indicated amounts of agonist in induction buffer. cAMP levels alterations were measured using a luminescence-based assay. Each point represents three data points, the error bars represent SEM. Data analysis was performed with GraphPad Prism software using a “log(agonist) vs. response, Variable slope” equation.


 Image

(B) Antagonist dose response curve. CHO cells with stable expression of dopamine D2 receptor were incubated with forskolin, receptor agonist and indicated amounts of antagonist in induction buffer. cAMP levels alterations were measured using a luminescence-based assay. Each point represents three data points, the error bars represent SEM. Data analysis was performed with GraphPad Prism software using a “log(inhibitor) vs. response, Variable slope” equation.