The B-Bridge Cyclic AMP (cAMP) Chemiluminescent Direct Immunoassay (CLIA) kits are designed to quantitatively measure cAMP present in cell lysates, plasma, urine, saliva, tissue and culture media samples. The supplied Sample Diluent will lyse cells, stabilize cAMP and stop phosphodiesterase activity. A cAMP standard is provided to generate a standard curve for the assay. The supplied Plate Primer solution is added to the wells of a coated white microtiter plate, followed by standards or diluted samples. A cAMP-peroxidase conjugate is then added to the wells.
The binding reaction is initiated by the addition of a polyclonal antibody to cAMP. After a 2 hour incubation the plate is washed and chemiluminescent substrate is added. The substrate immediately reacts with the bound cAMP-peroxidase conjugate. The generated chemiluminescent glow signal is measured. The concentration of the cAMP in the sample is calculated, after making correction for the dilution.
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. Cyclic AMP is produced by the enzyme adenylate cyclase, and the enzyme is activated by the hormones glucagon and adrenaline and by G protein. cAMP decomposition into AMP is catalyzed by the enzyme phosphodiesterase. Other biological actions of cAMP include regulation of innate immune functioning, axon regeneration, cancer, and inflammation.
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