FTO (human intracellular) ELISA Kit

FTO, Fat mass-and obesity-associated gene, was discovered as a responsible gene causing the mouse ‘fused toes’ mutation. The predicted 502-amino acid Fto protein has a calculated molecular mass of 58 kD and contains an N-terminal bipartite nuclear localization signal. FTO is widely expressed in a variety of human tissues, with highest levels in brain and pancreatic islets. Bioinformatics analysis indicates that FTO shares sequence motifs with iron- and 2-oxoglutarate (2OG)-dependent oxygenases. The FTO (human) (IntraCellular) ELISA Kit is to be used for the in vitro quantitative determination of human FTO in cell lysates or cell-based assays (screening). This assay is a sandwich Enzyme Linked-Immunosorbent Assay (ELISA) for quantitative determination of human FTO in cells. A monoclonal antibody specific for FTO has been precoated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, FTO is recognized by the addition of a purified polyclonal antibody specific for FTO (Detection Antibody). After removal of excess polyclonal antibody, HRP conjugated anti-rabbit IgG (Detector) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450 nm after acidification and is directly proportional to the concentration of FTO in the samples. This ELISA is specific for the measurement of natural and recombinant human FTO. It does cross-react with human adiponectin, human RBP4, human Nampt, human vaspin, human progranulin, human resistin, human clusterin, human GPX3, human sirtuin 1, human IDO, human IL-33, human ANGPTL3, human ANGPTL4, human FGF21, mouse progranulin, mouse ANGPTL3, mouse leptin, rat Nampt. The assay range is 0.156 – 10 ng FTO/ml and a detection limit of 50 pg/ml (based on adding two standard deviations to the mean value of the (50) zero standards).