本试剂盒应用双抗体夹心酶标免疫分析法测定标本中肾上腺髓质素水平。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肾上腺髓质素抗原、生物素化的抗人肾上腺髓质素抗体、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肾上腺髓质素呈正相关。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 human adrenomedullin, ADM concentrations in cell culture supernates, serum and plasma.
Introduction
Adrenomedullin (ADM) is a peptide associated with pheochromocytoma. It was discovered in 1993. The human ADM gene is localized to a single locus on Chromosome 11 with 4 exons and 3 introns. The ADM gene initially codes for a 185-amino acid precursor peptide, that can be differentially excised to form a number of peptides, including an inactive 53-amino acid ADM, e PAMP, adrenotensin and ADM95-146. Mature human ADM is activated to form a 52 amino acid, 6-amino acid ring, that shares moderate structural similarity to the calcitonin family of regulatory peptides (calcitonin, CGRP and amylin). Circulating ADM consists of both amidated (mature) and the glycated form (inactive, with the latter comprising the major form (85%). The measured to have a plasma half-life of 22min, mean clearance rate of 274 mL/kg/min, and apparent volume of distribution of 880+/- 150 mL/kg. Mature ADM is metabolised via aminopeptidase action.
At present ADM is believed to function through the combination of a few combinations of the calitonin receptor like receptor (CL) and receptor activity-modifying proteins (RAMP) complexes, as well as CGRP receptors. It is worth noting that unlike the classical one ligand-one receptor notion of receptor signalling, the interaction of both CL and RAMP at the membrane, is required for ADM to mediate its action. The outcome of ADM stimulatation of its receptor is the cellular production of both cyclic AMP (cAMP) and nitric oxide production. Some may find the production of these inside the cell to be at odds, since often they have opposing effects, but as yet, the timing of these effects remains to be studied.
ADM was initially identified as a vasodilator, some have cited this as the most potent endogenous vasodilatory peptide found in the body. Differences in opinion regarding the ability of ADM to relax vascular tone arises from the differences in the model system used. Other effects of ADM include increasing the tolerance of cells to oxidative stress and hypoxic injury and angiogenesis. ADM is seen as a positive influence in diseases such as hypertension, myocardial infarction, chronic obstructive pulmonary disease and other cardiovascular diseases, whereas it can be seen as a negative factor in potentiating the potential of cancerous cells to extend their blood supply and cause cell proliferation.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A antibody specific for ADM has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ADM present is bound by the immobilized antibody. An enzyme-linked antibody specific for ADM 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 ADM bound in the initial step. The color development is stopped and the intensity of the color is measured.
