The tissue inhibitors of metalloproteinases(TIMPs) are natural inhibitors of the matrix metalloproteinases, a group of zinc-binding endopeptidases involved in the degradation of the extracellular matrix. The TIMP3 gene is expressed in many tissues, with highest expression in the placenta. TIMP3 encodes a potent angiogenesis inhibitor and is mutated in Sorsby fundus dystrophy, a macular degenerative disease with submacular choroidal neovascularization. TIMP3 gene is mapped to 22q12.1-q13.2. Mutations in TIMP3 cause the autosomal dominant disorder Sorsby's fundus dystrophy(SFD).
Sample Type
Serum, cell culture supernates, saliva, plasma (heparin or EDTA).
Intended Use
Sandwich High Sensitivity BioAssay™ ELISA kit for Quantitative Detection of Equine TIMP-3.
Range
156pg/ml-10,000pg/ml
Specificity
Recognizes natural and recombinant Equine TIMP-3. There is no detectable cross-reactivity with other relevant proteins.
Test Principle
This Equine TIMP-3 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A monoclonal antibody from mouse specific for Equine TIMP-3 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO; Immunogen sequence: C24-P211) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Equine TIMP-3 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 Equine TIMP-3 amount of sample captured in plate.
Kit Components
96-well plate precoated with anti-Equine TIMP3 antibody, 1x plate Lyophilized recombinant Equine TIMP3 standard, 2x10ng/tube Biotinylated anti-Equine TIMP3 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer
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.
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