Description
Introduction: HRG (histidine-rich glycoprotein) contains two cystatin-like domains and is located in plasma and platelets. The physiological function is not yet known. It binds heme, dyes and divalent metal ions. It can inhibit rosette formation and interacts with heparin, thrombospondin, and the lysine-binding site of plasminogen. A potential prothrombotic effect of HRG is indicated by the inhibition of fibrinolysis and the reduction of inhibition of coagulation. Mutations in HRG lead to thrombophilia due to abnormal histidine-rich glycoprotein levels.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to HRG. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for HRG and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain HRG, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of HRG in the samples is then determined by comparing the O.D. of the samples to the standard curve