G protein-coupled receptors (GPCRs) are a large protein family of receptors that sense molecules outside the cell and activate inside signal transduction pathways and cellular responses. There are two principal pathways: cAMP and phosphatidylinositol.
cAMP Accumulation Assay
Creative BioMart offers cAMP Accumulation Assay for measuring cyclic AMP. Creative BioMart has developed the platform optimized for the detection of a specific range of cAMP concentrations, to enable assay sensitivity fine tuning. It gives high quality, cell-based assay screening in 96-, 384- or 1536-well formats .
- Cell-based functional assay
- Second messenger accumulation assay
- Assay formats 100 μL and 20 μL
- Miniaturization down to < 10μl
- Compatible with fresh or frozen cells
- Gs or Gi coupled receptor activation
- Assess agonists, antagonists, inverse agonists and allosteric modulators
- Phosphodiesterase (PDE) monitoring
- Adenylate cyclase activity assessment
Detection of cAMP is based on the competition between cAMP produced by cells and a biotinylated cAMP probe that is recognized by the streptavidin-Donor and anti-cAMP conjugated Acceptor beads. The beads are brought into proximity and a signal is detected. Increased intracellular concentrations of cAMP following Gs coupled GPCR activation by an agonist results in the displacement of the biotinylated cAMP probe and leads to a proportional signal decrease. The effect of antagonists and reverse agonists can similarly be detected. Gi coupled receptor activation can be detected after prestimulating cells with forskolin.