Background

This rat urokinase plasminogen activator activity assay is for the quantitative determination of active plasminogen activator in rat plasma.
Urokinase plasminogen activator is a serine that activates plasminogen to plasmin in the blood fibrinolytic system. It is also implicated in events related to cell invasion/migration (3).

Assay Principles

Functionally active uPA will bind to the biotinylated human PAI-1 coated on the microtiter plate. Only free active enzyme will react with the PAI-1 on the plate. Inactive or complexed enzyme will not be detected. After appropriate washing steps, monoclonal mouse anti-rat uPA primary antibody binds to the captured enzyme. Excess antibody is washed away, and bound monoclonal antibody is then reacted with the secondary antibody conjugated to horseradish peroxidase. TMB substrate is used for color development at 450 nm. A standard calibration curve is prepared along with the samples to be measured using dilutions of uPA.

Typical Standard Curve

References

1. Declerck, P.J., et al.; (1995) Thromb Haemostas: 74(5): 1305-9
2. Singh, I., et al.; (2003) Circulation: 107(6): 869-875
3. Kjøller, L,; (2002) Biol. Chem.: 383: 5-19
4. Yang, Y.H., et al.; (2001) J. Immunol.: 167(2): 1047-52