Description
Intended Uses: This Free Protein-S ELISA is a solid phase enzyme immunoassay for the determination of free Pro-tein-S in citrated human plasma.
Principle of the Assay: The Free Protein S is a sandwich ELISA using microplates coated with a capture antibody specific for human free Protein-S. Diluted plasma samples (1:51) are incubated in the wells allowing free Protein-S present in the plasma to bind to the antibody. The unbound fraction is removed by washing. After-wards anti-human Protein-S detection antibody conjugated to horseradish peroxidase (conjugate) is incubated and reacts with the antigen-antibody complex on the microwell surface. Following incuba-tion, unbound conjugate is washed off. Addition of TMB-substrate generates an enzymatic colorimetric (blue) reaction, which is stopped by diluted acid (color changes to yellow). The rate of color formation from the chromogen is measured in optical density units with a spectrophotometer at 450 nm. Using a curve prepared from the Reference Plasma provided with the kit, the free Protein-S antigen relative percent concentration in the plasma samples can be determined