Leptin, which is a product of ob gene, is a protein consisting of 167 amino acids and it is secreted from white adipose tissue. It is known that leptin acts on hypothalamus to decrease food intake and to reduce body weight, body fat, blood sugar and blood insulin in a healthy and an ob/ob mouse. Further, gene expression of neuropeptide Y (NPY) is suppressed by leptin. 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 of normal group.
The level well correlate with body fat and these observations show clearly that leptin concentration in human plasma reflects the tissue fat weight. The measurement of plasma leptin may be an excellent index of obesity. Although rat leptin shows a high homology (96%) with mouse leptin, it is observed that substitution of several amino acid residues occurs at both end N- and C- terminal region between human and rat leptin. These findings have required urgently to develop highly sensitive immunoassay system specific to rat leptin.
Yanaihara Institute Inc. already has rat leptin ELISA kit (YK050), and now we have developed the high sensitive (HS) enzyme immunoassay kit YK051, more sensitive than YK050 which has upgrade sensitivity to 39.1 pg/mL, for measuring rat leptin in its serum and plasma.
This ELISA kit is used for quantitative determination of rat leptin in its serum and plasma samples. The kit is characterized for sensitive quantification, high specificity and no influences with other components in samples and needlessness of sample pretreatment. Rat leptin standard is recombinant product.
This kit shows 0.41% cross reactivity to human leptin, 0.1% to mouse leptin, and it has no cross reactivity with rat IL-1α, IL-1β, rat TNF-α, human TNF-α and other cytokines.
This kit for determination of rat leptin in serum and plasma samples is based on two steps sandwich enzyme immunoassay. The 96-wells plate is coated with anti rat leptin monoclonal antibody. Rat leptin standard or samples and specific antibody are added to the wells for immunoreaction. During this immunoreaction, monoclonal antibody-antigen-specific antibody complex are formed on the surface of the wells. After incubation and plate washing, HRP-labeled antibody (goat anti rabbit IgG-HRP conjugated) is added to bind to specific antibody. Finally, HRP enzyme activity is determined by o-Phenylenediamine dihydrochloride (OPD) and the concentration of rat leptin is calculated.
|1. Antibody coated plate||MTP*1||1 plate
|Anti rat leptin monoclonal antibody|
|2. Standard antigen||lyophilized||11 vial
|Recombinant rat leptin (20ng)|
|3. Specific antibody||lyophilized||1 bottle||Rabbit anti rat leptin antibody|
|4. HRP labeled antibody||liquid||1 bottle
|Goat anti rabbit IgG-HRP conjugated|
|5. Substrate buffer||liquid||1 bottle
|0.015% Hydrogen peroxide|
|6. OPD tablet||ablet||2 tablets||o-Phenylenediamine dihydrochloride|
|7. Stopping solution||liquid||1 bottle
|8. Buffer solution||Liquid||1 bottle
|9. Washing solution
|10. Adhesive foil||4 pieces|
Leptin, ELISA (High Sensitivity)