Bio-immuno assay for the measurement of tPA activity. Assay designed with an ELISA plate coated with murine monocolonal antibody specific for tPA, then stabilized, which binds tPA present in plasma or in tested sample. tPA activity is then revealed through activation of plasminogen, in presence of tPA stimulator, and consequent cleavage of a substrate specific for Plasmin.
NTENDED USE:
Zymuphen tPA Activity kit is a bio-immunoassay for measuring tPA activity in human acidified citrated plasma or in purified milieu, using a microplate.
ASSAY PRINCIPLE:
In a first step, the diluted tested plasma or biological fluid is introduced into a microwell coated with a highly purified monoclonal antibody specific for human tPA, and that does not react with the tPA active site. When present, tPA is captured onto the solid phase. Following a washing step, in the presence of tPA stimulator (R2) that enhances the reaction, plasminogen (R2) is then converted into plasmin. Plasmin then reacts with its specific substrate (R1) and free pNA is released. tPA activity in the sample is directly proportional to the amount of free pNA measured at 405nm with a spectrophotometer.
Assay Characteristics
Kit Components
Total Assay Time:
about 2h
Detection Threshold:
≤ 1 IU/mL
Dynamic Range:
0-10 IU/mL
Intra-Assay CV:
≤ 8%
Inter-Assay CV:
≤ 10%
All the necessary for performing a 96 well microElisa plate. tPA is measured in acidified plasma for blocking tPA inhibitors.