Description:
Adipocytokines are a group of diverse effector molecules that are produced by adipose cells. Abnormal expression of adipokines such as Resistin is associated with the development of insulin resistance, diabetes and other metabolic and cardiovascular disorders in man. Mouse Resistin was described as a novel obesity-mediated adipocytokine that impairs glucose homeostasis by affecting both insulin-stimulated glucose uptake in adipose tissue and hepatic glucose production during fasting. However, there were initially two opposite views on human Resistin. Some studies have shown the ccorrelations between Resistin and obesity. Serum Resistin levels increase with increased adiposity and conversely, decline with decreased adiposity following medical treatment. However, other studies presented contradictory evidences that significantly decreased serum concentrations of resistin with increased adiposity. Resistin has also been shown to increase the expression of several pro-inflammatory cytokines including IL-1, IL-6, IL-12, and TNF-a via NF-kB.Principle:
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The ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay utilizes a mouse anti-human cytokine antibody for immobilization on the microtiter wells and goat anti-human cytokine antibodies along with streptavidin conjugated to horseradish peroxidase (HRP) for detection. The test sample is allowed to react simultaneously with the two antibodies, resulting in the molecules of the cytokine being sandwiched between the solid phase and enzyme-linked antibodies. After incubation, the wells are washed to remove unbound-labeled antibodies. A HRP substrate, TMB, is added to result in the development of a blue color. The color development is then stopped with the addition of Stop Solution changing the color to yellow. The concentration of the cytokine is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm. |
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