本试剂盒应用双抗体夹心酶标免疫分析法测定标本中TLR 9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TLR 9抗原、生物素化的抗人TLR 9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TLR 9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 human TLR 9 concentrations in cell culture supernates, serum, and plasma.
Introduction
Members of the Toll-like receptor (TLR) family are responsible for the recognition of pathogen-associated molecular patterns (PAMPs) expressed by a wide spectrum of infectious agents. To date, over thirteen TLRs have been reported in human and mouse. TLRs activate the NF-κB pathway, which regulates cytokine expression, through several adaptor molecules including MyD88, TIRAP/Mal and TRIF. Activation of the NF-κB pathway links innate and adaptive immune response by production of inflammatory cytokines such as IL-1, IL-6, IL-8, TNF-α, IL-12, chemokines and induction of costimulatory molecules such as CD80, CD86, and CD40. In addition to induction of the cytokine network, MyD88 binds FADD and triggers apoptosis through the Caspase cascade. Hence, activation of the apoptosis pathway via TLRs appears to contribute to the repertoire of defense mechanisms utilized by the innate immune response.
TLR 9, a receptor for CpG DNA, is localized mainly in endosomal/lysosomal compartments and has been implicated in the activation of autoreactive B cells in vitro.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for TLR 9 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TLR 9 present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for TLR 9 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 TLR 9 bound in the initial step. The color
development is stopped and the intensity of the color is measured.
