Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate NT5E/CD73 in samples. An antibody specific for NT5E/CD73 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyNT5E/CD73 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for NT5E/CD73 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 NT5E/CD73 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Ecto-5-prime-nucleotidase (5-prime-ribonucleotide phosphohydrolase; EC 3.1.3.5) catalyzes the conversion at neutral pH of purine 5-prime mononucleotides to nucleosides, the preferred substrate being AMP. The enzyme consists of a dimer of 2 identical 70-kD subunits bound by a glycosyl phosphatidyl inositol linkage to the external face of the plasma membrane.The enzyme is used as a marker of lymphocyte differentiation. Consequently, a deficiency of NT5 occurs in a variety of immunodeficiency diseases . Other forms of 5-prime nucleotidase exist in the cytoplasm and lysosomes and can be distinguished from ecto-NT5 by their substrate affinities, requirement for divalent magnesium ion, activation by ATP, and inhibition by inorganic phosphate
