Diapharma PAI-1 Activity is a bio-immuno assay for the measurement of PAI-1 activity.
Assay designed with an ELISA plate coated with active tPA, then stabilised, which binds active PAI-1 present in plasma or in tested sample. This latter, is then revealed with a specific murine monoclonal antibody, specific for PAI-1 and labelled with HorseRadish Peroxidase (HRP).
The kit contains a calibrator, referenced to the NIBSC international standard, and two control plasmas (low and high). This assay is specific for PAI-1 activity. Controls are included.
- COAT: Micro ELISA plate, containing 12 strips of 8 wells, coated with tPA, then stabilized; the plate is packed in an aluminium pouch hermetically sealed in presence of a desiccant.
- SD: 2 vials containing 50 ml of F-Sample Diluent, ready to use.
- Cal: 3 vials of PAI-1 Activity Calibrator, lyophilized. Each vial, when restored with 2 ml of F-Sample Diluent, allows obtaining the calibrator. The exact PAI-1 Activity concentration is indicated on the flyer provided in the kit (about 10 ng/ml).
- CI: 1 vial containing 1 ml of lyophilized Plasma PAI-1 Control I High (human plasma).
- CII: 1 vial containing 1 ml of lyophilized Plasma PAI-1 Control II Low (human plasma).
Note: The PAI-1 Activity and acceptancy ranges for controls I and II can vary from lot to lot, and are indicated on the flyer provided in the kit.
- IC: 3 vials of Anti-(h)-PAI-1-HRP immunoconjugate, a monoclonal antibody coupled to HRP, lyophilized.
- CD: 1 vial of 25 ml of Conjugate Diluent, ready to use.
- WS: 1 vial of 50 ml of 20 fold concentrated Wash Solution.
- TMB: 1 vial of 25 ml peroxidase substrate: 3,3′,5,5′ – Tetramethylbenzidine containing hydrogen peroxide. Ready to use.
- SA: 1 vial of 6 ml of 0.45M Sulfuric acid (Stop solution). Ready to use.
Note: Use only components from kits with the same lot number. Do not mix components from different lots of kits when running the assay.
The diluted tested plasma or biological fluid is introduced into a microwell coated with recombinant tPA. When present, PAI-1 binds to coated tPA. Only the active PAI-1 reacts with tPA and is fixed on the solid phase. Following a washing step, the immunoconjugate, which is a mouse monoclonal antibody specific for human PAI-1 and coupled to horse radish peroxidase (HRP), is introduced, and binds to its specific epitope on immobilized PAI-1. Following a new washing step, the peroxidase substrate, Tetramethylbenzidine (TMB) in presence of hydrogen peroxide (H2O2), is introduced and a blue color develops. The color turns yellow when the reaction is stopped with Sulfuric Acid. The amount of color developed is directly proportional to the amount of human PAI-1 Activity in the tested sample.