More info
Assay Range | 156 - 10,000 pg/mL |
Sensitivity | 10.0 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 Angiostatin K1-3 standard: 2 vials.
2. One 96-well plate coated with anti- Human Angiostatin K1-3 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
Angiostatin is an anti-angiogenic proteolytic fragment of Plasminogen. Plasminogen is produced primarily in the liver, but also in other tissues. Human Plasminogen contains an N-terminal activation peptide, five characteristically folded kringle domains, and a peptidase S1 domain. Cleavage of the activation peptide produces mature Plasminogen, which can be further cleaved between Arg580 and Val581 by tissue plasminogen activator (tPA) to produce the disulfide-linked two-subunit enzyme plasmin that plays a key role in breakdown of fibrin clots. Angiostatin was first identified as a protein consisting of kringles 1-4. Other anti-angiogenic forms include kringles 1-3 (K1-3), or 1-4 plus most of kringle 5 (K4.5). It is reported that K4.5 is the most active form. Angiostatin has exhibited potent anti-angiogenic and antitumor activity. It interferes with FGF2 and VEGF signaling, and inhibits endothelial cell proliferation and tube formation. It also inhibits endothelial cell and macrophage migration, and upregulates expression of antiangiogenic Thrombospondin-1 and macrophage IL-12. Angiostatin K1-3 is a non-glycosylated 259 amino acid polypeptide containing the first three kringle domains of plasminogen. Angiostatin K1-3 is a potent inhibitor of angiogenesis and tumor growth and shows increased inhibitory activity in comparison with kringles 1-4.