预期应用
ELISA法定量测定人血清、血浆或其它相关液体中透明质酸结合蛋白(HABP)含量。
介绍
体内存在多种可与HA 特异性结合的蛋白质,主要分布在基质和细胞膜上，人们将这些蛋白质统称为HA 结合蛋白(HA-binding proteins , HABP)。报道最多的基质内HABP 主要为软骨基质内的连接蛋白和蛋白聚糖,两者的核心蛋白均有与HA 特异性结合部位，识别HA 分子链中的十糖片段并与之结合。研究较为明确的细胞膜HABP(受体)主要有两类，CD44和RHAMM ( receptor for HA-mediated motility).CD44在不同细胞的表现形式可不同，具有众多的异构体。RHAMM是在成纤维细胞表面发现的，HA 介导的细胞移动就是通过该受体而产生的。
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HABP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HABP、生物素化的抗人HABP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HABP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 HA-bindingproteins,HABP concentrations in cell culture supernates, serum, and plasma.
Introduction
Hyaluronic Acid (HA), also called hyaluronate or hyaluronan, is a mucopolysaccharide widely distributed throughout the body. HA is produced mainly by fibroblasts and other specialized connective tissue cells.HABP is hyaluronic acid binding protein. Detection of 34 kDa HABP in the serum-free supernatant culture medium of fibroblasts was further evident by immunoblot analysis, thus confirming the secretory nature of HABP and its occurrence in the extracellular matrix.HABP binding to C32 cells and erythrocytes was independent of peptide charge or peptide structure. HABP efficiently prevented the basic fibroblast growth factor/epidermal growth factor (bFGF/EGF)-dependent proliferation of human umbilical vein endothelial cells.It also demonstrates that ITI-related proteins might be considered as HA-binding proteins.HABP staining revealed a stronger reactivity in the follicular keratinocytes surrounding the mucin-accumulated areas compared to the interfollicular keratinocytes. It was reported that human fibroblast 34-kDa hyaluronic acid-binding protein (HABP) is identical with P32, the protein co-purified with splicing factor SF-2.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for HABP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HABP present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for HABP 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 HABP bound in the initial step. The color development is stopped and the intensity of the color is measured.
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