cAMP(Cyclic adenosine monophosphate) ELISA Kit (E-EL-0056)

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal cAMP. During the reaction, Universal cAMP in the sample or standard competes with a fixed amount of Universal cAMP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal cAMP. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Universal cAMP in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Cyclic adenosine monophosphate (cAMP) is a universal second messenger present in virtually all cells. It was the first second messenger to be identified and is formed by intramolecular cyclization of ATP, being synthesized by adenylate cyclase and degraded to 5'-AMP by phosphodiesterases, so its intracellular concentration rises and falls rapidly after stimulation, converting extracellular signals into intracellular responses. Downstream of G protein-coupled receptors, cAMP regulates metabolic enzymes, ion transport, gene transcription and cell survival. It acts through three principal classes of effectors: protein kinase A, cyclic nucleotide-gated ion channels and the exchange protein directly activated by cAMP (Epac). Because these effectors are distributed differently among cell types, a single messenger produces distinct responses according to cellular context. Measurement of cAMP is widely used to assess G protein-coupled receptor pathway activity and the influence of drugs on cell function. Aberrant cAMP signaling is implicated in endocrine disorders, heart failure and certain cancers. Quantitative determination in cells, tissue and biological fluids supports receptor ligand screening and evaluation of adenylate cyclase or phosphodiesterase modulators, providing a readout of signal network status.