| 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. Rabbit 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. The Insulin-like Growth Factor 1 (IGF-1) BioAssay(TM) ELISA Kit is a sandwich ELISA for quantitative detection of rabbit IGF-1 in cell culture supernates, cell lysates, serum and plasma (heparin, EDTA). Detection Range: 156-10,000pg/ml Sensitivity: <50pg/ml Specificity: Recognizes natural and recombinant rabbit IGF-1. There is no detectable cross-reactivity or interference between IGF-1 and its analogs. Kit Components: 396698A: Microtiter Strips, 1x96 wells 396698B: IGF-1 Standard, 2x10ng 396698C: Anti-rabbit IGF-1 (Biotin), 1x100ul (1:100 dilution) USB Cat No Kit Component 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x100ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml 143702: Wash Buffer, 25X, 1x20ml Storage and Stability: Store *396698B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add samples and standards and incubate the plate at 37C for 90 minutes. Do not wash. 2. Add biotinylated antibodies and incubate the plate at 37C for 60 minutes. Wash plate 3 times with 0.01M TBS or PBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5 times with 0.01M TBS or PBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 25-30 minutes. 5. Add TMB stop solution and read. |
