CyclicAMP ELISA Kit

CAMP is a second messenger, used for intracellular signal transduction, such as transferring into cells the effects of hormones like glucagon and adrenaline, which cannot pass through the cell membrane. It is involved in the activation of protein kinases and regulates the effects of adrenaline and glucagon. cAMP also binds to and regulates the function of ion channels such as the HCN channels and a few other cyclic nucleotide-binding proteins such as Epac1. cAMP is synthesised from ATP by adenylyl cyclase located on the inner side of the plasma membrane. Adenylyl cyclase is activated by a range of signaling molecules through the activation of adenylyl cyclase stimulatory G (Gs) -protein-coupled receptors and inhibited by agonists of adenylyl cyclase inhibitory G (Gi) -protein-coupled receptors. Liver adenylyl cyclase responds more strongly to glucagon, and muscle adenylyl cyclase responds more strongly to adrenaline. cAMP decomposition into AMP is catalyzed by the enzyme phosphodiesterase.

Aviva Systems Biology cyclicAMP ELISA Kit (OKEH02525) is based on a competitive enzyme immunoassay technique. The microtiter well-plate in this kit has been pre-coated with an anti-cyclicAMP antibody. Sample or standards are added to the wells along with a fixed quantity of biotinylated cyclicAMP and incubated. The cyclicAMP found in the sample or standards competes with the biotinylated cyclicAMP for limited binding sites on the immobilized anti- cyclicAMP antibody. Excess unbound biotinylated cyclicAMP and sample or standard cyclicAMP is washed from the plate. Avidin-HRP conjugate is added, incubated and washed. An enzymatic reaction is then produced through the addition of TMB substrate which is catalyzed by the immobilized HRP to generate a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration is measured by reading the absorbance at 450 nm which is quantitatively proportional to the amount of biotinylated cyclicAMP captured in the well and inversely proportional to the amount of cyclicAMP which was contained in the sample or standard.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol