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 human vitronectin antigen in plasma, serum, culture media or tissue extract samples is easily performed with this 96 well strip format ELISA kit. Using this assay, values for plasma vitronectin in normal individuals were determined to be 127.5 +/- 13.1 ug/ml. Vascular samples from the saphenous vein, mammary artery, and human adipose tissue were also assayed for vitronectin, with values ranging from 5-40 ug/mg. The assay measures total vitronectin in the 0.5-100 ng/ml range. Samples giving human vitronectin levels above 100 ng/ml should be diluted in blocking buffer before use. Human vitronectin will bind to the capture antibody coated on the microtiter plate. This kit does not cross react with mouse vitronectin.After appropriate washing steps, anti-human vitronectin primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with the secondary antibody 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 human vitronectin in the samples. A standard calibration curve is prepared in blocking buffer using dilutions of purified human 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, Human Vitronectin Antigen Capture Plate, Secondary Antibody