本试剂盒应用双抗体夹心酶标免疫分析法测定标本中皮质醇水平。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皮质醇、生物素化的抗大鼠皮质醇抗体、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皮质醇呈正相关。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 Cortisol concentrations in cell culture supernates, serum, and plasma.
Introduction
Cortisol is the major glucocorticoid produced and secreted by the adrenal cortex. It affects the metabolism of protein, fat and carbohydrates; the maintenance of muscle and myocardial integrity; and the suppression of inflammatory and allergic activities. Production of cortisol from the adrenal cortex is dependent upon corticotrophin (ACTH), which is secreted by the anterior pituitary. The corticotrophin releasing factor (CRF) which regulates ACTH is secreted by the hypothalamus and is responsive to cortisol levels. Physical, psychological and surgical stress and diurnal variation will affect the rate of cortisol production. Corticosteroid-binding globulin and albumin bind approximately 90% of the cortisol secreted by the adrenal cortex. Bound cortisol circulates in an available but temporarily inactive state. The physiological activity of cortisol depends upon levels of the small fraction of circulating unbound cortisol.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for Cortisol has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Cortisol present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for Cortisol 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 Cortisol bound in the initial step. The color
development is stopped and the intensity of the color is measured.
