本试剂盒应用双抗体夹心酶标免疫分析法测定标本中α-GST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α-GST抗原、生物素化的抗大鼠α-GST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α-GST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 α-glutathione S-transferases, α-GST concentrations in cell culture supernates, serum, and plasma.
Introduction
Alpha-Glutathione S-transferase (aGST) is readily released from hepatocytes of both the centri-lobular and peri-portal regions in response to injury. In particular, the centri-lobular hepatocytes are highly susceptible to damage in various clinical conditions including allograft rejection, hepato-toxicity, as well as viral and chronic hepatitis. The rapid release of aGST into the circulation and its immediate removal qualifies this parameter as an advanced biomarker for the integrity of the liver parenchyma and liver status. In the liver alpha aGST is located in hepatocytes whereas pGST is confined to the intrahepatic bile duct cells. The heterogeneous GST subclass distribution suggests that the isoenzymes have unique in vivo functions in different hepatic regions and that the detection of GST subclass levels in biological fluids would be of significant use in monitoring the integrity of specific hepatic regions. aGST is unique to hepatocytes, and comprises 5 % of the soluble protein. Upon injury of hepatocytes aGST is readily released into the pericellular environment.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for aGST has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any aGST present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for aGST 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 aGST bound in the initial step. The color
development is stopped and the intensity of the color is measured.
