ELISA法定量测定人血清、血浆或其它相关液体中亲环蛋白A(CypA)含量。

 

本试剂盒应用双抗体夹心酶标免疫分析法测定标本中CypA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CypA抗原、生物素化的抗人CypA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CypA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。

 

This immunoassay kit allows for the specific measurement of human cyclophilin A ,CypA concentrations in cell culture supernates, serum, and plasma.

 

Cyclophilins, abbreviated CyP (not to be confused with CYP, which is the approved abbreviation for cytochrome P450), are proteins that bind to ciclosporin (cyclosporine A), an immunosuppressant which is usually used to suppress rejection after internal organ transplants. These proteins have peptidyl prolyl isomerase activity, which catalyzes the isomerization of peptide bonds from trans form to cis form at proline residues and facilitates protein folding.

Cyclophilin A, which exists in cytosol, has a beta barrel structure with two alpha helices and a beta-sheet. Other cyclophilins have similar structures to cyclophilin A. The cyclosporin A-cyclophilin A complex inhibits a calcium/calmodulin-dependent phosphatase, calcineurin, the inhibition of which is thought to suppress organ rejection.

 

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal

antibody specific for CypA has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CypA present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for CypA 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 CypA bound in the initial step. The color development is stopped and the intensity of the color is measured.