预期应用
ELISA法定量测定人血清、血浆、细胞培养物上清或其它相关液体中Rantes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Rantes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Rantes抗原、生物素化的抗人Rantes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Rantes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 Rantes concentrations in cell culture supernates, serum, and plasma.
Introduction
RANTES was identified in 1988, as a result of a subtractive hybridization screen for mRNA expressed by T cells, but not by B cells. In addition to T cells, RANTES is also produced by epithelial cells, monocytes, fibroblasts, and mesanglial cells. Platelets and eosinophils release RANTES from intracellular storage granules upon activation. Several high affinity receptors for RANTES have been described, including CCR1, CCR3, CCR5, and cytomegalovirus receptor US28. RANTES also binds to the erythrocyte Duffy antigen receptor (DARC) with low affinity. RANTES regulates the trafficking of cells expressing these receptors, including T cells (especially memory T cells), eosinophils, basophils, monocytes, macrophages, microglia, NK cells, dendritic cells, mast cells, and neurons. RANTES is also observed to induce Ca2+ flux and changes in protein tyrosine phosphorylation.
RANTES is important in a number of disease states, including inflammation, rheumatoid arthritis, experimental autoimmune encephalopathy, and anti-GMB glomerulonephritis. By recruiting eosinophils, RANTES plays a role in asthma and allergy, and RANTES antagonists capable of blocking eosinophil chemotaxis are currently under investigation as potential therapeutics. In humans, RANTES has an important role in HIV infection: R5 (macrophage tropic) HIV-1 uses CCR5 as a coreceptor, and several CCR5 ligands, including RANTES, MIP-1α, and MIP-1β, have been found to suppress HIV infection.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for Rantes has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Rantes present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for Rantes 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 Rantes bound in the initial step. The color development is stopped and the intensity of the color is measured.
|
