预期应用
ELISA法定量测定犬血清、血浆、细胞培养物上清或其它相关液体中Histamine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Histamine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Histamine抗原、生物素化的抗犬Histamine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Histamine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 canine Histamine concentrations in cell culture supernates, serum, and plasma.
Introduction
Histamine (ß-imidazolethylamine) is the most important mediator and is mostly found in the initial phase of an anaphylactic reaction ("immediate type" allergy). Histamine is developed by the enzymatic decarboxylation of histidine. In the organism, histamine is present in nearly all tissues, and it is mainly stored in the metachromatic granula of mast cells and the basophilic leukocytes. It is present in an inactive bound form and is only released as required. Histamine acts predominantly on smooth muscle and blood vessels.It is responsible for the bronchoconstriction occurring during the acute phase. In the vessels, its constrictive effect is limited to the venula, whereas arterioles are dilated. Furthermore, histamine causes a contraction of the cells of the vascular endothelium and increases the vascular permeability, thereby allowing higher-molecular substances to escape into the tissue.
Like several other mediators, histamine does not only mediate various clinical symptoms of anaphylaxis but also induces a series of effects which are directed towards a termination of the anaphylactic reaction. Histamine may inhibit the release of lysosomal enzymes from polymorphonoculear leukocytes, the degranulation of mast cells and basophiles and the production of complement components through mononuclear phagocytes. Furthermore, histamine can activate suppressor T cells and, thus, may inhibit the production of IgE. The biological action of histamine in tissue is guaranteed by three different surface receptors, i.e. H1, H2 and H3 receptors.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Histamine has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Histamine present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for Histamine 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 Histamine bound in the initial step. The color development is stopped and the intensity of the color is measured.
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