Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate ACAT1 in samples. An antibody specific for ACAT1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyACAT1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for ACAT1 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 ACAT1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:ACAT1 is a mitochondrially localized enzyme that catalyzes the reversible formation of acetoacetyl-CoA from two molecules of acetyl-CoA. Defects in this gene are associated with 3-ketothiolase deficiency, an inborn error of isoleucine catabolism characterized by urinary excretion of 2-methyl-3-hydroxybutyric acid, 2-methylacetoacetic acid, tiglylglycine, and butanone.The 427-amino acid precursor had a molecular mass of 45.2 kD. The sequence included a 33-residue leader peptide and a 394-amino acid subunit of the mature enzyme, which had a molecular mass of 41.4 kD. In all 4 cell lines, the T2 mRNA had the same 1.7-kb transcript as that of the control; however, content was reduced in 2 cell lines and normal in the other 2. Human T2 is a homotetramer of 41-kD subunits.
