Leptin, which is a product of ob gene, is a protein consisting of 146 amino acids and it is secreted from white adipose tissue. It is known that leptin acts on hypothalamus to decrease food intake and reduce body weight, body fat, blood sugar and blood insulin in a healthy and an ob/ob mice. Further, gene expression of neuropeptide Y (NPY) is suppressed by this protein. Recently, radioimmunoassay for leptin determination in human plasma has become available and leptin level in human patient group with obesity was found to increase in comparison with that in normal group. The plasma level well correlated with body fat and these observations showed clearly that leptin concentration in human plasma reflects tissue fat weight. The measurement of leptin in plasma or serum may be a good index of obesity. Although mouse leptin shows a high homology (96%) with rat leptin in amino acid sequences, it is observed that substitution of quite a number of residues occurs between human and mouse leptin. These findings have urged us to develop highly sensitive immunoassay system specific to rat or mouse leptin. Yanaihara Institute Inc. already has rat leptin ELISA kit (YK050 and YK051), and now we have developed mouse leptin ELISA kit for measuring mouse leptin in its serum. It will be a specifically useful tool for leptin researches.
This ELISA kit is used for quantitative determination of mouse leptin in its serum samples. The kit is characterized by its sensitive quantification and high specificity. In addition, it is not influenced by other constituents in samples. Standard antigen, mouse leptin, in this kit is a recombinant product.
This ELISA kit is highly specific to mouse leptin. It shows 33.8 % crossreactivity to rat leptin and no crossreactivity to human leptin.
This ELISA kit for determination of mouse leptin is based on a sandwich enzyme immunoassay. To the wells of plate coated with highly purified antibody against mouse leptin, standard antigen or sample is added for the 1st step immunoreaction. After the 1st step incubation and plate washing, biotinylated rabbit anti mouse leptin antibody is added as the 2nd step reaction to form leptin antibody- antigen -biotinylated leptin antibody complex on the surface of the wells. After the 2nd step incubation and rinsing out excess biotinylated antibody, horse radish peroxidase (HRP) labeled streptoavidin (SA) is added for binding to biotinylated leptin antibody. Finally, HRP enzyme activity is determined by 3,3’,5,5’-tetramethylbenzidine (TMB) and the concentration of mouse leptin is calculated.
Leptin, ELISA Kit