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预期应用
ELISA法定量测定大鼠血清、血浆、细胞培养物上清、组织匀浆或其它相关液体中cAMP含量。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定标本中cAMP水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入cAMP抗原、生物素化的抗大鼠cAMP抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的cAMP呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
This immunoassay kit allows for the specific measurement of rat cAMP concentrations in cell culture supernates, tissue homogenates, serum, and plasma.
Introduction
Adenosine 3’,5’-cyclic monophosphate (cAMP) is a ubiquitous second messenger involved in various cellular activities in many cell and tissue types. It is converted from adenosine triphosphate (ATP) via adenylyl cyclases (AC). There are at least 10 AC isoforms found in mammals, 9 of which are multi-transmembrane domain proteins (AC1 - AC9), and one soluble form has been described (sAC). Membrane subtypes may be activated by the stimulatory G protein a subunit (Gsa). Some examples of G protein-coupled receptors known to activate this pathway include those for the ligands epinephrine, dopamine, PGE2, adenosine, and glucagon. In contrast, the Gia subclass suppresses the activity of ACs . The intrinsic GTPase activity of the Gs subtype is classically suppressed by cholera toxin, while the Gi/o subtypes are inhibited by pertussis toxin. Ca2+ and its downstream effectors are also regulators of AC activity. Depending on the AC subtype, this regulation might be activating or inhibitory. Similarly, specific AC isoforms appear to be positively or negative regulated by Gbg subunits. Most isoforms are also potently activated exogenously by forskolin. sAC is structurally and biochemically distinct from membrane ACs. It is insensitive to forskolin and may be activated by bicarbonate. A large family of cyclic nucleotide phosphodiesterases regulates the levels of cAMP and guanosine 3’,5’-cyclic monophosphate (cGMP).
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for CAMP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CAMP present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for CAMP is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells
and color develops in proportion to the amount of CAMP bound in the initial step. The color
development is stopped and the intensity of the color is measured. | 
