Description
This is an Enzyme Immunoassay for the quantitative determination of total Prothrombin/Thrombin antigen in mouse serum, plasma, cell culture supernatant and tissue extracts. This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for Mouse Prothrombin has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any Prothrombin/Thrombin antigen present is bound by the immobilized antibody. Biotinylated primary antibody is then added and bound to the captured proteins. After washing away any unbound substances, streptavidinconjugated HRP is added and incubate. After washing steps are carried out, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Prothrombin/Thrombin bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450 nm ±2 nm. The concentration of Prothrombin/Thrombin in the sample is then determined by comparing the O.D of samples to the standard curve