| Insulin-like growth factor 1(IGF-1) that was once called somatomedin C, is a polypeptide protein hormone similar in molecular structure to insulin. It plays an important role in childhood growth and continues to have anabolic effects in adults. Equine IGF1 is a single chain 70aa polypeptide cross-linked by 3 disulfide bridges, with a calculated molecular mass of 7.6 kD. The IGF1 gene, mapped on 12q22-q24. contains 5 exons. Exons 1-4 encode the 195aa precursor(IGF1B), and exons 1, 2, 3, and 5 encode the 153-residue peptide(IGF1A). The structure of IGF1 resembles that of IGF2. And the IGF1 and IGF2 genes have complex structures with multiple promoters. The expression of both genes is regulated at the levels of transcription, RNA processing, and translation. IGF-1 is produced primarily by the liver as an endocrine hormone as well as in target tissues in a paracrine/autocrine fashion. Moreover, approximately 98% of IGF-1 is always bound to one of 6 binding proteins(IGF-BP). Furthermore, IGF-1 is one of the most potent natural activators of the AKT signaling pathway, a stimulator of cell growth and multiplication and a potent inhibitor of programmed cell death. Sample Type: Cell culture supernates, serum, plasma (heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Equine IGF-1. Sensitivity: <10pg/ml Range: 62.5pg/ml-4000pg/ml Specificity: Recognizes natural and recombinant Equine IGF-1. There is no detectable cross-reactivity with IGF-2. Test Principle: The Equine IGF-1 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A monoclonal antibody from mouse specific for Equine IGF-1 has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: G49-A118) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Equine IGF-1 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 IGF-1 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Equine IGF1 antibody, 1x plate Lyophilized recombinant Equine IGF1 standard, 2x10ng/tube Biotinylated anti-Equine IGF1 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. |
