Recombinant human urokinase — urokinase-type plasminogen activator (uPA / PLAU, EC 3.4.21.73) — expressed in E. coli, purified by ion-exchange and immunoaffinity chromatography and supplied lyophilised, 20 µg. An active serine protease that converts plasminogen to plasmin, the central trigger of pericellular proteolysis, fibrinolysis and matrix turnover. Use as an enzyme, as an antigen and immunisation standard, and as an affinity ligand — the matching antigen for the anti-urokinase antibodies cat. 168, cat. 335 and cat. 381, and the enzyme to pair with the uPA chromogenic substrate cat. 270.
SPECIFICATIONS
Catalogue number: 840 | Product: Urokinase (uPA / PLAU), human, recombinant
Source: recombinant, expressed in E. coli
Purity: 95% at 54 kDa by SDS-PAGE
Specific activity: 100 000 IU/mg — confirmed by lysis of a fibrin thin-layer gel (Astrup method) and by cleavage of a chromogenic uPA substrate
Format: lyophilised from a solution containing sodium phosphate 10 mM pH 7.5 and sodium chloride 150 mM
Purification: ion-exchange chromatography followed by affinity chromatography on immobilised anti-urokinase monoclonal antibody
Package size: 20 µg
STORAGE / RECONSTITUTION
Reconstitution: dissolve in distilled water.
Shipped at ambient temperature.
Lyophilised protein: store 2 years at −20 °C or lower.
Reconstituted protein: store at +4 °C for short-term use, or aliquot and freeze.
Avoid repeated freeze-thaw cycles of reconstituted material.
APPLICATIONS
- Active enzyme — plasminogen activator for fibrinolysis and plasmin-generation assays.
- Chromogenic activity assay — use with the uPA substrate pyroGlu-Gly-Arg-pNA cat. 270 to measure amidolytic activity and to screen inhibitors.
- Antigen / ELISA standard for uPA immunoassays — pair with cat. 168, cat. 335 or cat. 381 as capture or detection antibody.
- Immunogen for the production of anti-urokinase antibodies.
- Ligand for affinity chromatography (capture of uPA-binding partners such as uPAR and PAI-1).
- Western-blot positive control at ~54 kDa.
APPLICATION HIGHLIGHTS — HIGH-VALUE RESEARCH NICHES
- Tumour invasion and metastasis: the uPA / uPAR / PAI-1 axis is one of the best-validated prognostic systems in breast cancer, and active uPA is the reagent of record for invasion and Matrigel-transmigration assays.
- Inhibitor and drug screening: a defined, activity-specified enzyme (100 000 IU/mg) is the requirement for IC50 determination of small-molecule and antibody uPA inhibitors.
- Fibrinolysis and thrombolysis research: plasminogen activation, clot-lysis kinetics and fibrin-plate assays.
- Pericellular proteolysis and ECM remodelling: uPA-generated plasmin activates MMPs and liberates matrix-bound growth factors, linking it directly to collagen turnover.
- Recombinant, non-plasma source: an E. coli-expressed protein avoids the viral-screening and lot-variability constraints of urine- or plasma-derived urokinase.
- Neuroinflammation and wound repair: uPA is increasingly studied in blood-brain-barrier permeability, glial activation and re-epithelialisation.
