Cyclic AMP High-Sensitivity CLIA

$530.00

The Cyclic AMP High-Sensitivity CLIA is designed to quantitatively measure cAMP present in lysed cells, EDTA and heparin plasma, urine, saliva and tissue culture media samples.

SKU: SKT-208-96 Categories: , ,

Cyclic AMP High-Sensitivity CLIA

The Cyclic AMP High-Sensitivity CLIA is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.119 pmol/ml
Dynamic Range: Regular: 0.185 – 15 pmol/ml. Acetylated: 0.039 – 5 pmol/ml
Incubation Time: 2 hours
Sample Type: Cell lysates, EDTA Plasma, Heparin Plasma, Saliva, Tissue, Tissue Culture Media, Urine
Sample Size: 50 μL

Product manufactured in Canada by StressMarq.


Assay Principle

For tissue samples, saliva and urine, where the levels of cAMP are expected to be relatively high, the regular format for the assay can be used. For plasma samples and some dilute cell lysates an optional acetylation protocol can be used. This kit can measure as little as 1 femtomol cAMP per sample. The kit is unique in that all samples and standards are diluted into an acidic Sample Diluent, which contains special additives and stabilizers, for cAMP measurement. This allows plasma, urine and saliva samples to be read in an identical manner to lysed cells. Acidified samples of cAMP are stable and endogenous phosphodiesterases are inactivated in the Sample Diluent. A cAMP standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. A white microtiter plate coated with an antibody to capture sheep IgG is provided. Prior to the addition of any samples or standards a neutralizing Plate Primer solution is added to all the used wells. Standards or diluted samples, either with or without acetylation, are pipetted into the primed wells. A cAMP-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a sheep antibody to cAMP to each well. After a 2 hour incubation, the plate is washed and the chemiluminescent substrate is added. The substrate reacts with the bound cAMP-peroxidase conjugate to produce light The generated light is detected in a microtiter plate reader capable of reading luminescence. The concentration of the cAMP in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.


Related Products

Cyclic AMP EIA Kit
Cyclic GMP CLIA Kit (High-Sensitivity)
Cyclic GMP EIA Kit

Additional Information

Assay Background


Adenosine-3’,5’-cyclic monophosphate (cyclic AMP) is a second messenger that plays an important role in intracellular regulation. Specifically, it functions as a mediator of activity of several key hormones including epinephrine, glucagon, and ACTH (1-4). 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. cAMP decomposition into AMP is catalyzed by the enzyme phosphodiesterase. In the Human Metabolome Database there are 166 metabolic enzymes listed that convert cAMP (5). Other biological actions of cAMP include regulation of innate immune functioning (6), axon regeneration (7), cancer (8), and inflammation (9).

Typical Standard Curve


Manual

Product Manual