Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate 5-NT in samples. An antibody specific for 5-NT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any5-NT present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for 5-NT 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 5-NT bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:5'-nucleotidase (EC 3.1.3.5) is an enzyme which catalyzes the phosphorylytic cleavage of 5'nucleotides. Although originally found in snake venom, the activity of 5'nucleotidase has been described for bacteria and plant cells, and is widely distributed in vertebrate tissue. In mammalian cells the enzyme is predominantly located in the plasma membrane and its primary role is in the conversion of extracellular nucleotides (e.g. 5'-AMP), which are generally impermeable, to the corresponding nucleoside (e.g. adenosine) which can readily enter most cells. Consequently, the enzyme plays a key role in the metabolism of nucleotides.
