Mouse and Rat Leptin ELISA

Mouse and Rat Leptin
ELISA# of Samples: 1X96 Assays
Intended Use: The 40-055-200005 Mouse and Rat Leptin ELISA is a sandwich enzyme immunoassay for the quantitative measurement of mouse and/or rat leptin. Introduction: Leptin is a protein hormone with important effects in metabolism and regulating body weight. It is a single-chain 16 kDa protein consisting of 146 amino acid residues and encoded by the obese (ob) gene. Leptin is expressed predominantly by adipocytes small amounts of leptin are also secreted by cells in the epithelium of stomach and in the placenta. Leptin& acute; s effect on body weight is mediated through effects on hypothalamic centers where leptin receptors are highly expressed. Leptin has a dual action it decreases the appetite and increases energy consumption. A mutations in the ob gene of leptin or in the gene of leptin receptor causes hyperphagia reduced energy expenditure and severe obesity in the ob/ob mice. Ob gene knockout mice are also characterized by several metabolic abnormalities including hyperglucocorticoidemia hyperglycaemia hyperinsulinemia and insulin resistance. When ob/ob mice are treated with injections of leptin they lose their excess fat and return to normal body weight. Studies have shown that leptin appears to be a significant regulator of reproductive function. In addition leptin is involved in bone metabolism and plays a significant role as an immunomodulator. Areas of investigation:Energy metabolism and body weight regulationPrinciple of the test: In the Mouse and Rat Leptin ELISA Standards Quality Controls and samples are incubated in microplate wells pre-coated with anti-mouse leptin antibody. After 60 minutes incubation and washing biotin-labelled polyclonal anti-mouse leptin antibody is added to the wells and incubated with immobilized antibody-leptin complex for 60 minutes. After another washing streptavidin-HRP conjugate is added. After 30 minutes incubation and the last washing step the remaining conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured spectrophotometrically at 450 nm. The absorbance is proportional to the concentration of leptin. A standard curve is constructed by plotting absorbance values against concentrations of Standards and concentrations of unknown samples are determined using this standard curve. Storage and Stability: 250 ?l of each standard is sufficient to run the standard curve in duplicates. Reconstituted standards (4000 pg/ml) can be aliquoted and stored at -20 C until next use (up to 3 months). Avoid repeating freezing/thawing cycles.

May function as part of a signaling pathway that acts to regulate the size of the body fat depot. An increase in the level of LEP may act directly or indirectly on the CNS to inhibit food intake and/or regulate energy expenditure as part of a homeostatic mechanism to maintain constancy of the adipose mass.

Secreted.

Belongs to the leptin family.