cAMP Radioimmunoassay

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Details for:  cAMP Radioimmunoassay

Radio immunoassay for the quantitative determination of cAMP in human plasma and urine. The test can also be used for applications in other biological materials. For research use only. Not for use in clinical diagnostic procedures.

This assay is based on the principle of radioimmunoassays for cyclic nucleotides described by Steiner et al. (1972), modified by Harper and Brooker (1975). The kit includes two standard ranges. The less sensitive method (1.5-450 pmol/mL) is applicable for urine and other samples with high cAMP levels. The highly sensitive method (0.05-15 pmol/mL) includes a simple acetylation step and is suitable for plasma and tissue extracts.

In the cAMP radioimmunoassay the samples (or standards) are incubated over-night with 125I-labeled cAMP and the pre-precipitated antiserum. There is competition between the radioactive and the non-radioactive antigen for a fixed number of antibody binding sites. The amount of 125I-labeled antigen bound to the antibody is inversely proportional to the analysed concentration of the sample. When the system is in equilibrium, the reaction is stopped by the addition of a co-precipitating solution and a centrifugation step. The precipitate is counted in a gamma counter. Quantification of unknowns is achieved by comparing their activity with a response curve prepared using known standards. Because of its high specificity and sensitivity the assay is also suitable for the determination of cAMP in the adenylate cyclase assay in the presence of high concentrations of ATP.

Cyclic adenosin-3',5'-monophosphate (cAMP) serves as a second messenger in signal transduction of various hormones, like adrenaline, ACTH, LH, FSH, glucagon and calcitonin. The hormone itself is the first messenger, binding to specific membrane receptors of hormone-responsive cells. This leads to the activation of an enzyme located at the inner side of the cell membrane, called adenylate cyclase, which catalyses the synthesis of cAMP from ATP. The increased cellular level of cAMP initiates the activation of protein kinases.


For concrete data please consult the Instruction for Use in the download box on the right side.

Catalog No. RE11021

cAMP Radioimmunoassay
List Price $410
Kit size 100
Method RIA (DAK)
Incubation time 1 x 20 h
Standard range 0.05 - 15 pmol/mL acetylation, 1.5 - 450 pmol/mL without acetylation
Specimen / Volumes 20 µL acetyl. plasma, urine, cell and tissue suspension
Substrate / isotope 125I ≤ 150 kBq
Regulatory Status: For research use only

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