Human TIMP-3(Tissue Inhibitors of Metalloproteinase 3) ELISA Kit (E-EL-H1454)

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human TIMP-3. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TIMP-3 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human TIMP-3, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human TIMP-3. You can calculate the concentration of Human TIMP-3 in the samples by comparing the OD of the samples to the standard curve.

Tissue inhibitor of metalloproteinases 3 (TIMP-3) is a secreted glycoprotein of the TIMP family and the only member that is efficiently retained within the extracellular matrix after secretion, an unusual property conferred by high-affinity binding to heparan sulfate proteoglycans on cell surfaces and in the matrix. This localization focuses its inhibitory activity on pericellular proteolysis, and its expression is regulated during development and in response to tissue injury and inflammation. Through its N-terminal domain TIMP-3 binds the catalytic site of metalloproteinases in a tight 1:1 non-covalent complex, inhibiting a broad spectrum of matrix metalloproteinases as well as the sheddases ADAM10 and ADAM17. Because ADAM17 releases membrane-anchored ligands such as tumor necrosis factor alpha, inhibition by TIMP-3 attenuates cell-surface inflammatory and growth signaling, and the protein is additionally a potent inhibitor of angiogenesis. TIMP-3 behaves as a tumor suppressor and is frequently silenced by promoter methylation in cancer, while its dysregulation contributes to atherosclerosis and tissue-destructive remodeling. Heterozygous TIMP-3 mutations cause Sorsby fundus dystrophy, an inherited macular degenerative disease. Because the balance between metalloproteinases and this inhibitor determines whether matrix remodeling remains controlled, this ELISA kit enables quantitative determination of TIMP-3 in serum, plasma and tissue extracts, supporting studies of tumor, vascular and retinal disease.