The endothelins is a family of structurally and pharmacologically distinct peptides, which has been identified and sequenced in Canines. Three isoforms of Canine endothelin have been identified: endothelin-1, -2, and -3. Endothelin-1 is a potent, 21aa vasoconstrictor peptide produced by vascular endothelial cells. Endothelins are 21aa vasoconstricting peptides produced primarily in the endothelium having a key role in vascular homeostasis. Endothelins are implicated in vascular diseases of several organ systems, including the heart, general circulation and brain. Endothelins are proteins that constrict blood vessels and raise blood pressure. They are normally kept in balance by other mechanisms, but when they are over-expressed, they contribute to high blood pressure(hypertension) and heart disease. Sample Type: Cell culture supernates, serum, plasma (heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Canine Endothelin. Sensitivity: <0.5pg/ml Range: 3.9pg/ml-250pg/ml Specificity: Recognizes natural and recombinant Canine Endothelin. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Canine Endothelin BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Canine Endothelin has been precoated onto 96-well plates. Standards(Expression system for standard: human peptide, Immunogen sequence: C53-W73) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Canine Endothelin 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 Endothelin amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Canine EDN1 antibody, 1x plate Lyophilized recombinant Canine EDN1 standard, 2x1ng/tube Biotinylated anti-Canine EDN1 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 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.
