Mouse Prothrombin ELISA Kit
Prothrombin (aka Factor II) is a single-chain vitamin K dependent 579 amino acid glycoprotein zymogen. Prothrombin is proteolytically activated to thrombin by the prothrombinase enzyme complex in the coagulation cascade common pathway. The serine protease thrombin converts plasma fibrinogen to insoluble fibrin. Prothrombin levels are decreased by anticoagulant therapy, vitamin K deficiency and severe liver disease. Elevated plasma prothrombin is associated with a single nucleotide change at position 20210.
This mouse prothrombin assay is intended for the quantitative determination of prothrombin in mouse plasma, serum, tissue extracts, and cell culture media. The assay will not detect mouse thrombin. The assay measures prothrombin antigen in the 1-500 ng/ml range. If the unknown is thought to have high prothrombin levels, dilutions may be made in blocking buffer. A 1:1,000-1:5,000 dilution for normal mouse plasma is suggested for best results. Samples of mouse serum, tissue extracts and cell culture media may be applied directly to the plate. Strip well format. Reagents for up to 96 tests.
Mouse prothrombin will bind to the capture antibody coated on the microtiter plate. Thrombin and thrombin-antithrombin complex will not react with the antibody coated on the plate. After appropriate washing steps, biotinylated primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with horseradish peroxidase conjugated streptavidin. TMB substrate is used for color development at 450nm. A standard calibration curve is prepared along with the samples to be measured using dilutions of prothrombin. The amount of color development is directly proportional to the concentration of prothrombin in the sample.
Detection Target: Prothrombin
Range: 1 - 500 ng/ml
Sensitivity: 0.375 ng/ml
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