TIMP-2 gen is encoded by 5 exons spanning 83 kb of genomic DNA. TIMP-2 is 83 kilobase pairs (kb) long with exon-intron splicing sites located in preserved positions among the three members of the TIMP family. The gene for tissue inhibitor of metalloproteinases-2 is localized on human chromosome arm 17q25. TIMP-2 abrogates angiogenic factor-induced endothelial cell proliferation in vitro and angiogenesis in vivo independent of MMP inhibition. The standard product used in this kit is recombinant human TIMP-2 with the molecular mass of 22Kd and 194aa.
The Tissue Inhibitors of Metalloproteinase 2 (TIMP-2) BioAssay™ ELISA Kit is for the quantitative detection of rat TIMP-2 in cell culture supernates, serum and plasma (heparin, EDTA).
Detection Range
156-10000pg/ml
Specificity
Natural and recombinant rat TIMP-2
Test Principle
This rat TIMP-2 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for TIMP-2 has been precoated onto 96-well plates. Standards(NSO, C27-P220) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TIMP-2 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was 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 rat TIMP-2 amount of sample captured in plate.
Kit Components
353072A: Microtiter Strips, 1x96 wells *353072B: TIMP-2 Standard, 2x10ng 353072C: Anti-rat TIMP-2 (Biotin), 1x100ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x100ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml 143702: Wash Buffer, 25X, 1x20ml
Storage and Stability
Store *353072B powder at 4°C. Once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.