Cyclic AMP (cAMP) Direct Immunoassay kit is designed to quantitatively measure cAMP present in lysed cells, tissue, plasma, urine, saliva and culture media samples. The kit is unique in that all samples and standards are diluted into an acidic Sample Diluent which contains cAMP stabilizers and is designed to lyse cells. A cAMP standard is provided to generate a standard curve for the assay. A special Plate Primer is added to all the wells and standards or samples in Sample Diluent are pipetted into the primed clear microtiter plate. A cAMP-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of an antibody to cAMP. After 2 hours, the plate is washed and substrate is added. The substrate reacts with the bound cAMP-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is read. An optional Acetylation Format allows super low concentrations of cAMP to be measured.
Adenosine-3',5'-cyclic monophosphate, or cyclic AMP (cAMP), is one of the most important second messengers and a key intracellular regulator. Discovered by Sutherland and Rall in 1957, it functions as a mediator of activity for a number of hormones, including epinephrine, glucagon, and ACTH. Adenylate cyclase is activated by the hormones glucagon and adrenaline and by G protein. Liver adenylate cyclase responds more strongly to glucagon, and muscle adenylate cyclase responds more strongly to adrenaline. cAMP decomposition into AMP is catalyzed by the enzyme phosphodiesterase. In the Human Metabolome Database there are 166 metabolic enzymes listed that convert cAMP.
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