Description
Principle of the Assay: This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-TIMP-2 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TIMP-2 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-2 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TIMP-2 can be calculated.
Background: TIMP metallopeptidase inhibitor 2, also known as TIMP-2, is a 21-kD protein, which is secreted by human melanoma cells and binds to type IV collagenase proenzyme secreted by the same cells. The TIMP2 gene is a member of the TIMP gene family, It is encoded by 5 exons spanning 83 kb of genomic DNA. The proteins encoded by this gene family are natural inhibitors of the matrix metalloproteinases, a group of peptidases involved in degradation of the extracellular matrix. It has an amino acid sequence with many similarities to human tissue inhibitor of metalloproteinase (TIMP1). TIMP2 is a tissue inhibitor of metalloproteinases, TIMP2 induced a decrease in total protein tyrosine phosphatase (PTP) activity associated with beta-1 integrin subunits as well as dissociation of the phosphatase SHP1 from beta-1