本试剂盒应用双抗体夹心酶标免疫分析法测定标本中小鼠E-Selectin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小鼠E-Selectin、生物素化的抗小鼠E-Selectin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小鼠E-Selectin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 mouse E-selectin concentrations in serum and plasma.
Introduction
E-selectin (also known as CD62E and ELAM-1), P-selectin and L-selectin are members of a
small family of leukocyte adhesion molecules that share common structural motifs .Mature mouse E-selectin is an inducible, 117 kDa, type I transmembrane glycoprotein that is transiently and principally expressed on endothelial cells (EC) after activation by cytokines or bacterial endotoxin . Its extracellular region contains a 100 amino acid (aa) N-terminal C-type lectin domain, a 40 aa EGF-like motif, and six 60 aa short concensus repeats (SCRs,also known as Sushi domains). The molecule also has a 22 aa transmembrane segment and a short 33 aa cytoplasmic region that can transduce a signal in endothelial cells via mitogen-activated protein (MAP) kinase . E-selectin from different species contains varyingnumbers of SCRs .
E-selectin mediates the initial rolling and subsequent stable adhesion of leukocytes, and allows
for leukocyte activation by chemokines, leading to their extravasation at sites of inflammation. In general, E-selectin binds sialyl Lewis-x displayed on a number of glycoproteins or proteoglycans, including E-selectin ligand (ESL), P-selectin glycoprotein ligand-1 (PSGL-1) , cutaneous lymphocyte antigen (CLA; a variant of PSGL-1) , L-selectin (2) and 2-integrin . The role for many of these potential
E-selectin ligands is not clearly understood. Integrin activation may involve E-selectin, either
indirectly as a consequence of E-selectin binding to leukocyte E-selectin ligand(s), or through
direct integrin-E-selectin interaction. The binding of E-selectin to the leukocyte integrin
CD11/CD18 can lead to integrin activation.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for E-SELECTIN has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any E-SELECTIN present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for E-SELECTIN 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 E-SELECTIN bound in the initial step. The color development is stopped and the intensity of the color is measured.
