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 Mouse Angiopoietin-2 standard: 2 vials.
2. One 96-well plate precoated with anti- Mouse Angiopoietin-2 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
Angiopoietin-2 (Ang-2) is a member of the vascular growth factor family that also includes Ang-1 and Ang-4 in humans. Ang-2 is characterized by an N-terminal coiled-coil domain and a C-terminal fibrinogen-like domain. Ang-2 is approximately 60% identical to its homolog Ang-1. Mature mouse Angiopoietin-2 shares 86% and 96% amino acid (aa) sequence identity with human and rat Angiopoietin-2, respectively. Expression of human Ang-2 is restricted postnatally to pro-angiogenic regions including the placenta, ovaries, and uterus, while mouse Ang-2 is widely expressed in the mouse embryo.
Both Ang-1 and Ang-2 are ligands for the endothelial cell receptor tyrosine kinase Tie-2. The regulation of Ang-2/Tie-2 signaling is complicated, leading to inhibition in certain cell types or conditions, and activation in others. It is reported that Ang-1, but not Ang-2, could stimulate Tie-2 tyrosine phosphorylation in human umbilical vein endothelial cells (HUVECs), suggesting that Ang-2 could act as a competitive inhibitor of Ang-1 signaling. Overexpression of Ang-2 leads to disruption of embryonic blood vessel formation, a phenotype similar to that of Tie-2 knockouts, supporting further that Ang-2 acts as an Ang-1/Tie-2 inhibitor. In contrast, altering incubation times or elevating Ang-2 concentrations can result in Tie-2 activation in HUVECs. Ang-2 promotes endothelial cell death and vessel regression by itself, but can promote new vessel formation together with VEGF. Ang-2-mediated destabilization of mature vessel structures in the presence of VEGF may play a role in the angiogenesis of tumor tissues. It is shown that Ang-2 is elevated in colon carcinoma, gastric carcinoma, glioblastoma, glioma, hepatocellular carcinoma, neuroblastoma and non-small cell lung cancer.