Product Name: Vitronectin EIA Kit
# of Samples: 1X96 Assays
Intended Use: The Vitronectin EIA Kit is an in vitro enzyme immunoassay (EIA) kit for quantitative determination of human vitronectin (VN) in plasma, serum, cultured cell extracts, cell culture supernatants, and other biological fluids.
This kit is for research use only. It is not for use in diagnostic or therapeutic procedures.
Introduction: The adhesion protein vitronectin (VN), or serum spreading factor (identical to complement S-protein) is a major cell adhesion protein in plasma(1-7), usually present at the concentration of 200 - 300 mg/l. VN is also present in the extracellular matrix of tissues, and atherosclerotic plaques(4,5,8). VN contributes in various ways to the regulation of the immune and hemostatic systems, in particular at the blood-endothelium interphase(9). These functions include its role as complement inhibitor(10) and as ?scavenger protein? for reaction products of the blood coagulation cascade such as the thrombin-antithrombin III complex(11-13). Moreover, VN constitutes the major binding and stabilizing protein for plasminogen activator inhibitor-1(14). Through binding to different cytoadhesion receptors, termed integrins(15), VN may directly interact with endothelial cells(16) and platelets(17) and thereby localize its diverse function at certain pericellular sites. In liver disorders with high fibroblastic activity such as liver cirrhosis, a significant diminution of plasma vitronectin levels was observed(18,19).
Principle of the test: The VN EIA Kit is a solid phase EIA based on a sandwich method that utilizes two mouse monoclonal anti-VN antibodies to detect VN by one-step procedure. One of the antibodies has been precoated, and blocked against non-specific biuding. Then, samples, standards and peroxidase (POD)-labelled anti-VN antibody are simultaneously added to the wells of plates. During the incubation, VN is bound to anti-VN (solid phase) on one side and tagged by POD-anti-VN on the other. The reaction between POD and substrate (H2O2, and tetramethylbengidine) results in colour development with intensities proportional to the amount of VN present in samples and standards. The amount of VN can be quantitated by measuring the absorbance using an EIA plate reader. Accurate sample concentrations of VN can be determined by comparing their specific absorbances with those obtained for the standards plotted on a standard curve.
Storage and Stability: This kit is shipped at 2 - 8C and should be stored at 2 - 8C if not used. Under this condition, the kit is stable until expiration date on label.
Sensitivity and Specificity: This kit specifically measures VN with no detectable cross reaction with human fibronectin, laminin, collagen type I, or collagen type III. This kit cannot be used to measure mouse VN. The application of this kit for quantitating VN from other sources has not been tested.
Product Name: Vitronectin EIA Kit
Function: Vitronectin is a cell adhesion and spreading factor found in serum and tissues. Vitronectin interact with glycosaminoglycans and proteoglycans. Is recognized by certain members of the integrin family and serves as a cell-to-substrate adhesion molecule. Inhibitor of the membrane-damaging effect of the terminal cytolytic complement pathway.
Function: Somatomedin-B is a growth hormone-dependent serum factor with protease-inhibiting activity.
Subunit: Exists in two forms: a single chain 75 kDa form (V75) and a clipped form composed of two chains (65 kDa and 10 kDa) (V65+V10) which are held together by a disulfide bond. Interacts with SERPINE1/PAI1 and insulin.
Subcellular Location: Secreted, extracellular space.
Tissue Specificity: Plasma.
Domain: The SMB domain mediates interaction with SERPINE1/PAI1. The heparin-binding domain mediates interaction with insulin.
Ptm: Sulfated on 2 tyrosine residues.
Ptm: N- and O-glycosylated (By similarity).
Ptm: Phosphorylation on Thr-69 and Thr-76 favors cell adhesion and spreading.
Ptm: It has been suggested that the active SMB domain may be permitted considerable disulfide bond heterogeneity or variability, thus two alternate disulfide patterns based on 3D structures are described with 1 disulfide bond conserved in both.
Similarity: Contains 4 hemopexin-like domains.
Similarity: Contains 1 SMB (somatomedin-B) domain.