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Monoclonal Anti-cAMP Antibody Based Direct cAMP EIA Kit without Acetylation

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生化科技->原料供應->特用化學

Monoclonal Anti-cAMP Antibody Based Direct cAMP EIA Kit without Acetylation

產品規格

Model No:	80203

Country	US
Brand	NewEast
Size	kit
Minimum Order	1

產品描述

Adenosine 3', 5'-cyclic monophosphate (cyclic AMP; cAMP) modulates various physiological functions such as cardiovascular biology, learning and memory, olfaction, immune response, asthma and kidney function (1,2). cAMP is produced from ATP by adenylyl cyclases and is degraded by phosphodiesterases. Stimulation of adenylyl cyclases or inhibition of phosphodiesterases can increase cellular cAMP concentrations. Blockers of adenylyl cyclase-activating receptors and inhibitors of the cAMP-specific phosphodiesterases are used for treating human diseases. For example, blocking agents for cAMP-increasing beta-adrenergic receptors (beta-blockers) are used for treating abnormal heart rhythms, high blood pressure (hypertension), myocardial infarction and heart failure. Inhibitors of cAMP specific phosphodiesterase types 2 and 4 are being tested for cognition enhancement.

To screen for inhibitors or stimulators of cellular cAMP levels, it is essential to have a sensitive, selective and reproducible method to measure the cAMP concentrations. This is especially true for the initial screenings given the possible weaker effects of larger pools of compounds.

Currently available other ELISA kits measuring cAMP levels are based on the non-affinity-purified polyclonal anti-cAMP antibody. Despite the claimed selectivity, these polyclonal anti-cAMP antibodies display certain cross-reactivity with ATP. Given that ATP is the substrate for the cAMP production, it is very desirable to have an antibody with high specificity towards cAMP over ATP.

NewEast Biosciences cAMP ELISA kit is based on the unique mouse monoclonal anti-cAMP antibody. This monoclonal anti-cAMP antibody displays >108 fold of selectivity over ATP, cGMP, and other nucleoside analogues. NewEast Biosciences cAMP ELISA kit provides significantly improved sensitivity and selectivity over other kits based on polyclonal anti-cAMP antibodies. Our monoclonal anti-cAMP antibody-based ELISA kit also avoids the batch-to-batch variations associated with polyclonal antibody productions from animals, thus providing the reproducibility in the long run.

Furthermore, while polyclonal anti-cAMP antibodies used in other ELISA kits have higher affinity for acetylated cAMP than non-acetylated cAMP, NewEast Biosciences monoclonal anti-cAMP antibody has similar affinities to non-acetylated and acetylated cAMP molecules. Therefore, acetylation treatments of samples and standards are not needed in NewEast Biosciences cAMP ELISA kit. This significantly reduces the time for the assay. The avoidance of organic reagents used in the acetylation process provides a safe and healthy work environment.

Principle Outline
NewEast Biosciences cAMP ELISA Kit is a competitive immunoassay to measure the cAMP levels, either from cell extracts or from in vitro adenylyl cyclase assays. Briefly, multi-well plates are coated with goat-anti-mouse serum. cAMP in cell extracts or in in vitro adenylyl cyclase assays will competitively bind to the monoclonal anti-cAMP antibody in the presence of fixed amounts of cAMP-conjugated horse-radish peroxidase or alkaline phosphatase. Known amounts of cAMP are used as standards to generate the calculation curve. After a short incubation, the excess reagents are washed away and substrate is added. The multiwell plates are then read on a microplate reader at 450 nm or 405 nm. The intensity of the yellow color is inversely proportional to the concentration of cAMP in samples. The measured optical density is used to calculate the concentration of cAMP in samples based on the curve from the cAMP standards.

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