Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PAH in samples. An antibody specific for PAH has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPAH present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PAH 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 PAH bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Phenylalanine hydroxylase is the rate-limiting enzyme of the metabolic pathway which degrades excess phenylalanine. The other substrates in the reaction are molecular oxygen and tetrahydrobiopterin. Tetrahydrobiopterin is a member of the group of redox biochemicals known as pteridines.
PAH is the gene that encodes for phenylalanine hydroxylase. It was the research on phenylalanine hydroxylase by Seymour Kaufman that led to the discovery of tetrahydrobiopterin as a biological cofactor. Phenylalanine hydroxylase is a tetramer composed of four monomers, that is, composed of 4 identical subunits. Each subunit is in turn composed of three domains, a regulatory domain, a catalytic domain, and a tetramerization domain
