Principle of the Assay: AT3Ab ELISA kit applies the competitive enzyme immunoassay technique utilizing a polyclonal anti-AT3Ab antibody and an AT3Ab-HRP conjugate. The assay sample and buffer are incubated together with AT3Ab-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the AT3Ab concentration since AT3Ab from samples and AT3Ab-HRP conjugate compete for the anti-AT3Ab antibody binding site. Since the number of sites is limited, as more sites are occupied by AT3Ab from the sample, fewer sites are left to bind AT3Ab-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The AT3Ab concentration in each sample is interpolated from this standard curve.
NCBI and Uniprot Product Information
Uniprot Description
SERPINC1: Most important serine protease inhibitor in plasma that regulates the blood coagulation cascade. AT-III inhibits thrombin, matriptase-3/TMPRSS7, as well as factors IXa, Xa and XIa. Its inhibitory activity is greatly enhanced in the presence of heparin. Defects in SERPINC1 are the cause of antithrombin III deficiency (AT3D). AT3D is an important risk factor for hereditary thrombophilia, a hemostatic disorder characterized by a tendency to recurrent thrombosis. AT3D is classified into 4 types. Type I: characterized by a 50% decrease in antigenic and functional levels. Type II: has defects affecting the thrombin- binding domain. Type III: alteration of the heparin-binding domain. Plasma AT-III antigen levels are normal in type II and III. Type IV: consists of miscellaneous group of unclassifiable mutations. Belongs to the serpin family.
Protein type: Inhibitor; Secreted; Secreted, signal peptide
Cellular Component: extracellular space; extracellular region
Molecular Function: serine-type endopeptidase inhibitor activity; heparin binding; protease binding; protease inhibitor activity
Biological Process: hemostasis; negative regulation of inflammatory response; negative regulation of peptidase activity; blood coagulation; response to nutrient