Intended use 
This immunoassay kit allows for the in vitro quantitative determination of General cyclicAMP concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids. 
Test principle 
The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General cyclicAMP, During the reaction, General cyclicAMP in the sample or standard competes with a fixed amount of biotin-labeled General cyclicAMP for sites on a pre-coated Monoclonal antibody specific to General. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General cyclicAMP in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cyclic adenosine monophosphate and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1:4	1:8	1:16
serum(n=5)	85-94%	82-91%	110-119%	101-111%
EDTA plasma(n=5)	96-106%	101-110%	100-112%	100-110%
heparin plasma(n=5)	90-99%	103-114%	114-123%	106-116%

 

General Annotation

 

Function:

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.

 

Location:

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.

Sequence:DRVYIHPFHL 

 

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