Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate Tie-2 in samples. An antibody specific for Tie-2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyTie-2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for Tie-2 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of Tie-2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Tie-2 is a receptor tyrosine kinase that is expressed primarily on endothelial cells (ECs) and plays a critical role in vascular development. The Tie-2 gene encodes a protein of 1122 amino acids. The extracellular region has three distinct structural motifs including two immunoglobulin (Ig)-like loops separated by three EGF-like repeats, and three repeats with fibronectin type III homology located after the second Ig loop. The intracellular portion of Tie-2 contains two tyrosine kinase domains that, when phosphorylated, interact with a number of binding partners including Grb2, Grb7, Grb14, Shp2, the p85 subunit of phosphatidylinositol 3-kinase (PI3K), and Dok-R. Deletion of the last 16 amino acids of the intracellular C-terminus results in increased levels of autophosphorylation, suggesting that it may play an autoinhibitory role.
