Description
Principle of the Assay: This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-TIMP-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TIMP-1 polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were 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 TIMP-1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TIMP-1 can be calculated.
Background: Tissue inhibitor of metalloproteinase 1 (TIMP1), also known as Erythroid Potentiating Activity (EPA), is a 28,000 MW glycoprotein that is expressed from the several tissues of organisms. This protein is a member of the TIMP family. The glycoprotein is a natural inhibitor of the matrix metalloproteinases (MMPs), a group of peptidases involved in degradation of the extracellular matrix. It plays a major role in modulating the activity of interstitial collagenase as well as a number of connective tissue metalloendoproteases. Decreased TIMP1 and increased cathepsin V/L2 levels might play a role in the matrix degradation that is a hallmark of keratoconus corneas. It is able to promote cell proliferation in a wide range of cell types, and may also have an anti-apoptotic function