Vitronectin is an abundant plasma glycoprotein that helps regulate coagulation, fibrinolysis, complement activation, and cell adhesion. Vitronectin binds to glycosaminoglycans, collagen, plasminogen and urokinase receptors. It also may control the clearance of vascular thrombi by binding and stablilizing PAI-1. In binding PAI-1, it extends the lifetime of active PAI-1. Vitronectin may also be involved in the regulation of bone metabolism. The sensitive quantitative measurement of total mouse vitronectin antigen in plasma, serum, culture media or tissue extract samples is easily performed with this 96 well strip format ELISA kit. The vitronectin concentration in mouse serum has been estimated at 300 ug/ml by semi-quantitative immunoblotting. The assay measures total vitronectin in the 0.1-100 ng/ml range. Samples giving mouse vitronectin levels above 100 ng/ml should be diluted in blocking buffer before use. A 1:10,00 to 1:40,000 dilution for mouse plasma or serum is suggested for best results. Mouse vitronectin will bind to the capture antibody coated on the microtiter plate. After appropriate washing steps, biotin labeled anti-mouse vitronectin primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with streptavidin conjugated to HRP. Following an additional washing step, TMB substrate is used for color development at 450 nm. Color development is proportional to the concentration of mouse vitronectin in the samples. A standard calibration curve is prepared in blocking buffer using dilutions of purified mouse vitronectin and is measured along with the test samples. All reagents and standards are provided in these ELISA kits.
Suggested additional reagents: 10X Wash Buffer, TMB Substrate, Mouse Vitronectin Antigen Capture Plate, Avidin-HRP