Neurotrophin-3(NT-3) is a new member of the nerve growth factor gene family, which plays an important role in the development and maintenance of the vertebrate nervous system.NT-3 and its receptor, called neurotrophic tyrosine kinase receptor type 3(Ntrk3, also called TrkC), are expressed early and throughout embryogenesis. NT-3 is one of five neurotrophin growth factors which shape the development of the nervous system by regulating neuronal survival and differentiation. NT-3 may be one of the central nervous system-derived factors that mediate neural crest(NC) cell proliferation in vivo. NT-3 has been mapped to Canine chromosome 12p and Canine chromosome 6. Sample Type: Cell culture supernates, serum. Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Canine Neurotrophin-3. Sensitivity: <2pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Canine Neurotrophin-3. There is cross-reactivity with NT-4 (<2.5%), and no detectable cross- reactivity with any other cytokine. Test Principle: The Canine Neurotrophin-3 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for Canine Neurotrophin-3 has been precoated onto 96-well plates. Standards(Expression system for standard: sf21, Immunogen sequence: Y139-T257) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Canine Neurotrophin-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 Canine Neurotrophin-3 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Canine Neurotrophin-3 antibody, 1x plate Lyophilized recombinant Canine Neurotrophin-3 standard, 2x10ng/tube Biotinylated anti-Canine Neurotrophin-3, 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 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
