本试剂盒应用双抗体夹心酶标免疫分析法测定标本中CC16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CC16抗原、生物素化的抗大鼠CC16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CC16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 CC16 concentrations in cell culture supernates, serum, and plasma.
 
Introduction
Rat Clara Cell Protein (CC16, CC10 and also called uteroglobin, urinary protein 1 or Clara Cell Secretory Protein) belongs to the family of secretoglobins and is a secreted protein product of non-ciliated bronchiolar Clara cells. Its function remains to be elucidated but there is convincing data suggesting its phospholipase A2 inhibitory activity as well as a number of other immunomodulatory features including inhibition of interferon gamma signaling and Th1 vs. Th2 lymphocyte regulation. It was proposed as a potential peripheral marker of respiratory epithelial injury and bronchial dysfunction. Clara Cell Protein 16 concentrations have been determined in both serum and bronchoalveolar lavage fluid in numerous studies since 1994. In serum, its increase is associated with age, asbestos, nitrogen chloride and ozone exposure, sarcoidosis and high PEEP ventilation. Decreased serum CC16 levels are found after pulmonary resection, in silica-exposed workers, smokers and in asthma. Decreased CC16 concentrations were also found in the amniotic fluid of fetuses suffering from pulmonary hypoplasia caused by various mechanisms (diaphragmatic hernia, diabetic fetopathy, Turner and Down syndrome). In pleural effusions, the CC16 concentration appears to be associated with its diffusion from the lung as evidenced by high CC16 levels in cardiac pleural congestion. 
 
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for CC16 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CC16 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for CC16 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 CC16 bound in the initial step. The color development is stopped and the intensity of the color is measured.