预期应用

ELISA法定量测定兔血清、血浆、组织匀浆、细胞培养物上清或其它相关液体中IgA含量。

 

实验原理

本试剂盒应用双抗体夹心酶标免疫分析法测定标本中IgA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IgA、生物素化的抗兔IgA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IgA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 rabbit IgA concentrations in cell culture supernates, serum, tissue homogenates and plasma.

 

Introduction

Secretory IgA (IgA) consists of two IgA monomers joined by the J-chain and furthermore a secretory component. It is secreted in plasma cells based in the lamina propia of mucosal membranes. Synthesis of IgA is independent from the synthesis of serum IgA. This means lack of serum IgA does not necessarily mean a lack of IgA. Secretory IgA is the major immunoglobulin in saliva, tears, colostrum, nasal mucous, mother´s milk, tracheobronchial and gastrointestinal secretes. It plays a major role in preventing adherence of microorganisms to mucosal sites, in activation of the alternative complement pathway and in activating inflammatory reactions. Newborns are provided with IgA by mother´s milk and are passively immunized against gastrointestinal infections.

 

Test principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IgA has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IgA present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for IgA 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 IgA bound in the initial step. The color development is stopped and the intensity of the color is measured.