Human Factor V will bind to the capture monoclonal antibody coated on the microtiter plate. Factor V and Va will react with the antibody 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 human Factor V. Color development is proportional to the concentration of Factor V in the samples.
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