cAMP Direct Immunoassay Detection Kit (Fluorometric) provides a sensitive method for detecting adenylate cyclase activity in biochemical or cell-based assay systems. Compared to other ELISA cAMP assay kits, this kit eliminates the tedious acetylation step. The kit uses AbRed Indicator as a fluorimetric substrate to quantify the HRP activity. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation. The fluorescent product formed is proportional to the activity of HRP-cAMP conjugate. This kit is based on the competition between HRP-labeled cAMP and free cAMP for a fixed number of cAMP antibody binding sites. HRP-cAMP is displaced from the HRP-cAMP/anti-cAMP antibody complex by unlabeled free cAMP. In the absence of cAMP, HRP-cAMP conjugate is bound to anti-cAMP antibody exclusively. However, the unlabeled free cAMP in the test samples competes for anti-cAMP antibody with the HRP-cAMP antibody conjugate, therefore inhibits the binding of HRP-cAMP to anti-cAMP antibody.
Adenosine 3", 5" cyclic monophosphate (cAMP) is an important second messenger in intracellular signal transduction. Monitoring cAMP levels is one of the most common ways to screen for agonists and antagonists of GPCRs.
Functional Studies more details
Components: 1 x 96 tests; 10X Wash Solution: 1 x 10ml; AbRed Indicator: 1 vial; Anti-cAMP-coated 96-well plate: 1 unit; Assay Buffer: 1 x 20ml; cAMP Standard: 1 x 33µg; Cell Lysis Buffer: 1 x 10ml; HRP-cAMP Conjugate: 1 vial; Hydrogen Peroxide Solution: 1 x 50µl; Substrate Buffer: 1 x 10ml
Compatible Sample Types:
Urine, Plasma, Tissue, Adherent cells, Suspension cells, Cell culture media