Description
Principle of the assay: human Angiopoietin-2 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Angiopoietin-2 has been precoated onto 96-well plates. Standards(NSO,Y19-F496) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Angiopoietin-2 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Angiopoietin-2 amount of sample captured in plate.
Background: Angiopoietin 1 and Angiopoietin 2 are important for development of the endothelium, by regulating tyrosine phosphorylation of the membrane receptor Tie 2. Angiopoietin 2 is only 60% homologous with Angiopoietin 1. Angiopoietin-2 is a naturally occurring antagonist of angiopoietin-1 that competes for binding to the TIE2 receptor and blocks ANGPT1-induced TIE2 autophosphorylation. Angiopoietin 1 binding to Tie 2 causes phosphorylation of the receptor. Angiopoietin 2 competes for this binding, and thus blocks receptor phosphorylation. Angiopoietin 2 expression occurs at sites of vascular remodelling: dorsal aorta and major aortic branches, ovary, placenta and uterus