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AQP5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AQP5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AQP5抗原、生物素化的抗大鼠AQP5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AQP5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 Aquaporin 5 (AQP5) concentrations in cell culture supernates, serum, and plasma.

 

Introduction

Sidhaye et al. (2006) observed a dose-responsive decrease in Aqp5 abundance in mouse lung epithelial cells exposed to hypotonic medium. Hypotonic reduction of AQP5 was augmented and reduced, respectively, by conditions that activated or inhibited Trpv4 (605427). Hypotonic reduction of AQP5 required extracellular calcium and was associated with increased intracellular calcium. The response to hypotonicity was recapitulated by coexpression of TRPV4 and AQP5 in human embryonic kidney cells. Sidhaye et al. (2006) concluded that AQP5 abundance is tightly controlled along a spectrum of extracellular osmolalities and that its abundance in hypotonic conditions can be regulated by TRPV4 activation.

 

Test principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for AQP5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any AQP5 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for AQP5 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 AQP5 bound in the initial step. The color development is stopped and the intensity of the color is measured.