Cyclic AMP EIA Kit
The Cyclic AMP EIA Kit is for Research Use Only
Size: 1×96 wells
Sensitivity: 0.64 pmol/ml
Dynamic Range: Regular: 0.617 – 150 pmol/ml. Acetylated: 0.156 – 10 pmol/ml
Incubation Time: 2.5 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. 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 clear microtiter plate coated with an antibody to capture sheep IgG is provided and 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 substrate is added. The substrate reacts with the bound cAMP-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. 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.
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