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- Assay Type Competitive ELISA
- Sample Types Cell Lysates, Urine, Plasma, Saliva, Tissue Culture Media
- Sensitivity 0.64 pmol/mL (Regular), 0.083 pmol/mL (Acetylated)
- Species Cyclic AMP is identical across species
- Samples/Plate 40 in Duplicate (Regular), 39 in Duplicate (Acetylated)
- Assay Duration 2.5 Hours
- Readout Colorimetric, 450 nm
- Standard Curve
The Cyclic AMP Direct ELISA Kit quantitatively measures Cyclic AMP in cell lysates, urine, plasma, saliva, tissue, and tissue culture media. The Cyclic AMP Direct ELISA Kit is a competitive ELISA with a run time of 2.5 hours. This kit is multi-format and can detect both acetylated and non-acetylated cAMP in a variety of sample types. Please read the complete kit insert for more information before performing this assay.
Use our provided Cyclic AMP standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our donkey anti-sheep IgG antibody. Add the Cyclic AMP peroxidase conjugate and the Cyclic AMP sheep antibody. Then incubate the mixture covered for 2 hours, shaking at room temperature. The immunological reaction occurs between the anti-Cyclic AMP antibody, the Cyclic AMP in the sample or standard, and the Cyclic AMP-peroxidase conjugate. As the Cyclic AMP concentration in the sample increases, the bound Cyclic AMP-peroxidase conjugate decreases, causing a decrease in signal and vice versa.
After the 2-hour incubation, wash away the excess Cyclic AMP-peroxidase conjugate and add the TMB substrate. The TMB substrate reacts with the bound Cyclic AMP-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the Cyclic AMP concentration in the samples.
Adenosine-3’, 5’-cyclic monophosphate, or cyclic AMP (cAMP), is one of the most important second messengers and a critical intracellular regulator. Sutherland and Rall discovered it in 1957. Cyclic AMP mediates activity for several 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. The enzyme phosphodiesterase catalyzes cAMP decomposition into AMP. The Human Metabolome Database lists 166 metabolic enzymes that convert cAMP.