Description:

Tumour Necrosis Factor alpha (TNFa), is an inflammatory cytokine produced by macrophages/monocytes during acute inflammation and is responsible for a diverse range of signaling events within cells, leading to necrosis or apoptosis. The protein is also important for angiogenesis that is critical to the growth, progression, and metastasis of solid tumors. Furthermore, TNFa is associated with obesity. It is chronically elevated in adipose tissues of obese rodents and humans and may represent an important link between obesity and insulin resistance. In both obese mice and humans, TNFa is overexpressed in adipose tissue. TNFa inhibits insulin signaling, at least in part by blocking insulin receptor tyrosine kinase activity and inducing serine phosphorylation of insulin receptor substrate-1 (IRS-1). However, it is unclear what the physiological stimulator of TNF-a production by adipocyte during obesity is and how IRS-1 inhibits the tyrosine kinase activity of the insulin receptor after TNF-a treatment of the cells. A better understanding of the connection(s) between the TNF-a and the insulin signaling pathways could be important to find a cure for the state of insulin resistance observed during obesity.

Principle:

MouseCytokine ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay utilizes rabbit anti-mouse antibodies for immobilization on the microtiter wells and rabbit anti-mouse 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 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 VEGF is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

Data:

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