|
预期应用
ELISA法定量测定大鼠血清、血浆、细胞培养物上清或其它相关液体中iNOS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iNOS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iNOS抗原、生物素化的抗大鼠iNOS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iNOS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 iNOS concentrations in cell culture supernates, serum, and plasma.
Introduction
Nitric oxide (NO) is a short-lived molecule (t1/2 = seconds) capable of diffusing across membranes and reacting with a variety of targets. A reaction with O2 in aqueous solutions produces the relatively unreactive nitrate (NO3 -) and nitrite (NO2 -) ions as products. In the presence of superoxide (O2 -), however, NO reacts extremely rapidly to produce the very reactive and toxic peroxynitrite (ONOO-) which subsequently decomposes into additional highly reactive intermediates. NO ultimately exerts its biological effects by reacting either directly or through other reactive nitrogen intermediates with a variety of targets such as heme groups, Fe-S or Zn-S clusters, sulfhydryl groups or various other chemical substrates. This diversity of potential targets is reflected in the large number of different systems that utilize NO as a mediator and provides ample opportunity for abnormal regulation and development of pathological effects.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for INOS has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any INOS present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for INOS 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 INOS bound in the initial step. The color development is stopped and the intensity of the color is measured.
| 
