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.
Sample Type:Cell culture supernates, serum and plasma (heparin, EDTA)
Intended Use:For quantitative detection of rat TIMP-2 in cell culture supernates, serum and plasma (heparin, EDTA).
Sensitivity:<10pg/ml
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:96-well plate precoated with rat TIMP-2 antibody, 1Lyophilized recombinant rat TIMP-2 standard, 10ng/tube×2Biotinylated rat TIMP-2 antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB stop solution, 10ml
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
