Description
Description: This assay employs a two-site sandwich ELISA to quantitate HADH in samples. An antibody specific for HADH has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HADH present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for HADH 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 HADH bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: HADH is a member of the 3-hydroxyacyl-CoA dehydrogenase gene family. The encoded protein functions in the mitochondrial matrix to catalyze the oxidation of straight-chain 3-hydroxyacyl-CoAs as part of the beta-oxidation pathway. Its enzymatic activity is highest with medium-chain-length fatty acids. Mutations in this gene cause one form of familial hyperinsulinemic hypoglycemia. The human genome contains a related pseudogene.Human HADH encodes a deduced 314-amino acid protein composed of a 12-residue mitochondrial import signal peptide and a 302-residue mature HADH protein with a calculated molecular mass of 34.3 kD. The sequence of the mature protein shows 94% identity with HADH from pig heart. Northern blot analysis revealed expression of HADH in liver, kidney, pancreas, heart, and skeletal muscle