Assay Range | 62.5--4000 pg/mL |
Sensitivity | 5.0 pg/mL |
Size | 96T |
Storage | Store at 2 - 8ºC. Keep reconstituted standard and detection Ab at -20 ºC |
Assay Principle | Sandwich ELISA |
Sample volume | 100 µL final volume, dilution factor varies on samples |
Detection Method | Chromogenic |
Kit Components
1. Recombinant Human uPA standard: 2 vials
2. One 96-well plate precoated with anti-Human uPA Ab
3. Sample diluent buffer: 12 mL - 1
4. Detection antibody: 130 µL, dilution 1:100
5. Streptavidin-HRP: 130 µL, dilution 1:100
6. Antibody diluent buffer: 12 mL x1
7. Streptavidin-HRP diluent buffer: 12 mL x1
8. TMB developing agent: 10 mL x1
9. Stop solution: 10 mL x1.
10. Washing solution (20x): 25 mL x1.
Background
Urokinase-type Plasminogen Activator-1 (uPA), encoded by the PLAU gene in humans, is a secreted serine protease that plays an important role in extracellular matrix (ECM) remodeling, cellular migration, inflammation, and cancers. uPA is composed of an EGF-like growth factor domain (GFD), a Kringle domain (KD), a connecting peptide, and a peptidase domain. uPA is produced as an inactive zymogen that is cleavable to generate the active uPA which is a disulfide-linked dimer made up of the N-terminal A chain (EGF-like and Kringle domains) and the C-terminal B chain (peptidase domain). A secondary cleavage removes the N- terminal fragment (ATF) containing the EGF-like and Kringle domains produce a truncated form of uPA consisting of the active B chain disulfide-linked to a portion of the connecting peptide.
uPA is a ligand for the uPA receptor uPAR/CD87. The binding of uPA to uPAR induces recruitment of a second uPAR molecule to form a ternary complex. When bound to uPAR, uPA can cleave uPAR and remove the ligand-binding D1 domain of uPAR, leaving a 40 kDa uPAR fragment associated with the cell. uPA binding to uPAR promotes cell proliferation and migration by inducing the activation of PDGF R beta and EGF R either directly or through complexes with integrins. It is reported that levels of uPA and/or uPAR in the plasma, ascites, urinary, or cerebrospinal fluid are elevated in a variety of invasive and metastatic human cancers.