本试剂盒应用双抗体夹心酶标免疫分析法测定标本中Testosterone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Testosterone抗原、生物素化的抗人Testosterone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Testosterone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 Free-Testosterone concentrations in serum, plasma and cell culture supernates.
Introduction
Testosterone is formed in the Leydig Cell, then capillaries and veins carry it to the periphery or it traverses testicular myoid cells and enters the seminiferous tubules where it involved in spermatogenesis. In the seminiferous tubules testosterone stimulates primary spermatocytes to form secondary spermatocytes and finally young spermatocytes. Testicular secretion accounts for 95 % of the circulating testosterone present in men. In the female, ovary and adrenal secrete small amounts of testosterone,however the majority of testosterone in the blood derives from metabolism of androstenedione.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Testosterone has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Testosterone present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody specific for Testosterone 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 Testosterone bound in the initial step. The color development is stopped and the intensity of the color is measured.
