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MIP-1β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MIP-1β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MIP-1β抗原、生物素化的抗人MIP-1β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MIP-1β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 MIP-1β concentrations in cell culture supernates, serum, and plasma.

 

Introduction

The two variants of the Macrophage Inflammatory Protein 1, designated MIP-1α and MIP-1β, belong to the β or CC chemokine subfamily. The two chemokines were originally purified from lipopolysaccharidetreated murine monocytic cell lines, hence their designation as "Macrophage Inflammatory Protein". MIP-1α and MIP-1β are produced by stimulated leukocytes as well as other tissue cells and tumor cells. At least seven variants of human MIP-1β, corresponding to the products of independantly cloned cDNA, including Act-2, G-26, HC-21, HIMAP, H400, pAT 744 and MAD-5, have been described. At the protein levels, mature human MIP-1β has a length of 69 amino acids and shows a homology of 70% with MIP-1α. MIP-1β is a natural ligand for the chemokine receptors CCR3, CCR5 and CCR8.

The effect of MIP-1α and MIP-1β on monocytes and T cells include chemotaxis, a rise in intracellular [Ca2+], expression of integrins and increased adhesion to endothelial cells. The two chemokines provide an important signal for T cell activation resulting in enhanced proliferation, IL-2 secretion and cell surface IL-2 receptor expression. MIP-1α and MIP-1β chemoattract and degranulate NK cells and enhance NK cells as well as CTL mediated cytolysis. Both chemokines induce chemotaxis and [Ca2+] increase in dendritic cells.

 

Test principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal

antibody specific for MIP-1β has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MIP-1β present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for MIP-1β 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 MIP-1β bound in the initial step. The color development is stopped and the intensity of the color is measured.