Human Leptin ELISA

Human leptin is a protein of 167 amino acids that plays a key role in regulating energy intake and energy expenditure, including the regulation (decrease) of appetite and (increase) of metabolism. Leptin is produced by adipose tissue and the level of circulating leptin is directly proportional to the total amount of fat in the body. Once leptin has bound to the Ob-Rb receptor, it activates Stat3, which is phosphorylated and travels to the nucleus where it mediates gene expression. One of the main effects on gene expression is the down-regulation of the expression of endocannabinoids, responsible—among their many other functions—for increasing appetite. There are other intracellular pathways activated by leptin, but less is known about how they function in this system. Although leptin is a circulating signal that reduces appetite, in general, obese people who are resistant to the effects of leptin have an unusually high circulating concentration of leptin, in the same way that people with type 2 diabetes are resistant to the effects of insulin. Leptin is also found to stimulate endothelial cell proliferation and angiogenesis.

A. Benefits:

• Cost-effective analysis - High Quality with the most competitive price.

• Efficient and flexible - Multiple samples can be analyzed simultaneously with a flexible number of assay wells.

• Specific- A pair of highly selective antibodies specifically against human leptin.

• Simple procedure All in one system with a pre-coated 96-well plate.

B. Principle of the technology

Leptin ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay utilizes a monoclonal antibody against a distinct determinant on human leptin for immobilization on the microtiter wells and a mouse anti-leptin antibody conjugated to horseradish peroxidase (HRP) for detection. The test sample is allowed to react simultaneously with the two antibodies, resulting in the leptin molecules being sandwiched between the solid phase and enzyme-linked antibodies. After incubation, the wells are washed to remove any unbound-labeled antibodies. An 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 leptin is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

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