试剂盒利用竞争酶联免疫分析方法来测定细胞提取物或体外腺苷酸环化酶实验中的cAMP的水平。包被羊抗鼠多克隆抗体在酶标板上，细胞提取物或体外腺苷酸环化酶实验中的cAMP与固定数量的标记辣根过氧化物酶的cAMP竞争性的结合抗cAMP单克隆抗体，采用已知浓度的cAMP标准品来做标准曲线。检测450nm波长下的OD值，吸光度OD值与样品中cAMP的浓度呈反比关系。基于cAMP标准品所得曲线，通过测得的OD值可以计算出样品中cAMP的浓度，本试剂盒IC50（50%B/B0）约12pmol/mL，检测限约0.15 pmol/mL。Product Highlight
产品优势：
基于小鼠独特的抗cgmp单克隆抗体
市面上唯一含单克隆抗cgmp抗体的试剂盒(其他试剂盒均为多克隆抗cgmp抗体)
单克隆抗体对cAMP和GTP的选择性明显高于多克隆抗体
避免了与动物多克隆抗体生产相关的批间变异，从而提供了可重复性
ProductDescription
Adenosine 3′, 5′-cyclic monophosphate (cyclic AMP; cAMP) modulatesvarious physiological functions such as cardiovascular biology,learning and memory, olfaction, immune response, asthma and kidneyfunction (1,2). cAMP is produced from ATP by adenylyl cyclases andis degraded by phosphodiesterases. Stimulation of adenylyl cyclasesor inhibition of phosphodiesterases can increase cellular cAMPconcentrations. Blockers of adenylyl cyclase-activating receptorsand inhibitors of the cAMP-specific phosphodiesterases are used fortreating human diseases. For example, blocking agents forcAMP-increasing beta-adrenergic receptors (beta-blockers) are usedfor treating abnormal heart rhythms, high blood pressure(hypertension), myocardial infarction and heart failure. Inhibitorsof cAMP specific phosphodiesterase types 2 and 4 are being testedfor cognition enhancement.
To screen for inhibitors or stimulators of cellular cAMP levels,it is essential to have a sensitive, selective and reproduciblemethod to measure the cAMP concentrations. This is especially truefor the initial screenings given the possible weaker effects oflarger pools of compounds.
Currently available other ELISA kits measuring cAMP levels arebased on the non-affinity-purified polyclonal anti-cAMP antibody.Despite the claimed selectivity, these polyclonal anti-cAMPantibodies display certain cross-reactivity with ATP. Given thatATP is the substrate for the cAMP production, it is very desirableto have an antibody with high specificity towards cAMP overATP.
NewEast Biosciences cAMP ELISA kit is based on the unique mousemonoclonal anti-cAMP antibody. This monoclonal anti-cAMP antibodydisplays >108 foldof selectivity over ATP, cGMP, and other nucleoside analogues.NewEast Biosciences cAMP ELISA kit provides significantly improvedsensitivity and selectivity over other kits based on polyclonalanti-cAMP antibodies. Our monoclonal anti-cAMP antibody-based ELISAkit also avoids the batch-to-batch variations associated withpolyclonal antibody productions from animals, thus providing thereproducibility in the long run.
PrincipleOutline
NewEast Biosciences cAMP ELISA Kit is a competitive immunoassay tomeasure the cAMP levels, either from cell extracts or from in vitroadenylyl cyclase assays. Briefly, multi-well plates are coated withgoat-anti-mouse serum. cAMP in cell extracts or in in vitroadenylyl cyclase assays will competitively bind to the monoclonalanti-cAMP antibody in the presence of fixed amounts ofcAMP-conjugated horse-radish peroxidase or alkaline phosphatase.Known amounts of cAMP are used as standards to generate thecalculation curve. After a short incubation, the excess reagentsare washed away and substrate is added. The multiwell plates arethen read on a microplate reader at 450 nm or 405 nm. The intensityof the yellow color is inversely proportional to the concentrationof cAMP in samples. The measured optical density is used tocalculate the concentration of cAMP in samples based on the curvefrom the cAMP standards.
