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PF4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PF4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PF4抗原、生物素化的抗大鼠PF4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PF4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 Platelet Factor 4 (PF4) concentrations in cell culture supernates, serum, and plasma.

 

Introduction

Platelet Factor  (PF4) is a 70 amino acid, 30,000 D molecular ratio protein. PF4 is released from activated platelet α granules in a tetrameric form complexed with platelet proteoglycan. Upon release, the half-life of PF4 is very short, less than 5 minutes, as it quickly binds to endothelial cell glycosaminoglycans where it is stored. PF4 possesses a powerful anti-heparin activity as it binds to heparin, forming a stochiometric complex, where 1 mg of PF4 will inhibit 27 IU of heparin.

 

Test principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for PF4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PF4 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for PF4 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 PF4 bound in the initial step. The color development is stopped and the intensity of the color is measured.