IGF1R, also called IGFR or insulin-like growth factor 1 receptor, is a protein that found on the surface of human cells. This gene is mapped to 15q26.3. IGF1R belongs to the large class of tyrosine kinase receptors. This receptor mediates the effects of IGF-1, which is a polypeptide protein hormone similar in molecular structure to insulin. It plays an important role in growth and continues to have anabolic effects in adults - meaning that it can induce hypertrophy of skeletal muscle and other target tissues. IGF1R also plays a critical role in transformation events. It is highly overexpressed in most malignant tissues where it functions as an anti-apoptotic agent by enhancing cell survival. Intended Use: Sandwich ELISA quantitative detection of human IGF1R in cell culture supernates, serum and plasma (heparin, EDTA). Range: 156-10000pg/ml Sensitivity: <10pg/ml Specificity: Natural and recombinant human IGF1R Crossreactivity: There is no detectable cross-reactivity with other relevant proteins. Test Principle: This human IGF1R BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for IGF1R has been precoated onto 96-well plates. Standards (NS0, E31-N932) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IGF1R 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 human IGF1R amount of sample captured in plate. Kit Components: 352914A: Microtiter Strips, 1x96 wells *352914B: IGF1R Standard, 2x10ng 352914C: Anti-human IGF1R (Biotin), 1x130ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (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 Storage and Stability: Store *352914B 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 100ul of samples and standards and incubate the plate at 37C for 90 min. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 min. Wash plate 3 times with 0.01M PBS or TBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5 times with 0.01M PBS or TBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 25-30 minutes. 5. Add 100ul TMB stop solution and read.
