Obesity is a well-known risk factor of type 2 diabetes mellitus and is strongly associated with insulin resistance. Resistin (also called FIZZ3/ADSF) is an adipocyte-derived peptide first identified during a search for targets of thiazolidinediones. Serum concentrations of resistin are markedly increased in obese mice and are decreased by treatment with thiazolidinediones. It was also found that administration of an anti-resistin antibody increases insulin-stimulated glucose uptake in obese mice and that treatment of normal mice with recombinant resistin impairs insulin action. Thus, resistin might link obesity with insulin resistance and diabetes in mice models. However, subsequent studies in rodent models have produced disparate findings on the role of resistin in obesity and insulin resistance. In humans, while the expression of resistin in human adipocytes is very low compared with that seen in rodents and does not differ between normal, insulin-resistant or type 2 diabetic individuals, a more recent study using a large size of case suggests that the plasma resistin levels are increased in type 2 diabetes. Therefore determination of the plasma resistin levels may be important for understanding onsets of metabolic diseases such as type 2 diabetes or obesity. This assay is a sandwich ELISA for quantitative determination of mouse resistin in biological fluids. A monoclonal antibody specific for resistin 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, resistin is recognized by the addition of a biotinylated polyclonal antibody specific for resistin (Detection Antibody). After removal of excess biotinylated antibody, HRP labeled streptavidin (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 450nm after acidification and is directly proportional to the concentration of resistin in the samples. The assay range is 0.47-30ng/ml resistin/ml. The lowest level of resistin that can be detected by this assay is 100pg/ml.Sample Type:Cell and tissue culture supernatants, urine, plasma, serum, as well as many other biological fluids.Intended Use:For the quantitative determination of mouse Resistin in biological fluids.Sensitivity:100pg/mlRange:0.47-30ng/ml Resistin/mlSpecificity:MammalianTest Principle:This assay is a sandwich ELISA for quantitative determination of mouse resistin in biological fluids. A monoclonal antibody specific for resistin 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, resistin is recognized by the addition of a biotinylated polyclonal antibody specific for resistin (Detection Antibody). After removal of excess biotinylated antibody, HRP labeled streptavidin (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 450nm after acidification and is directly proportional to the concentration of resistin in the samples.Kit Components:Pre-coated Microtiter Plate: 12x8 well stripsWash Buffer (10X): 1x50mlDiluent (5X):1x50mlDetection Antibody: 1x12mlDetector 100X (HRP Labeled Streptavidin): 1x150ulMouse Resistin Standard (lyophilized): 16ngMouse Resistin QC Sample (lyophilized): 1 vialSubstrate Solution I (TMB): 1x6mlSubstrate Solution II (Peroxidase): 1x6mlStop Solution: 1x12mlStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.