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 surpressed 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 Insutitute Inc. has developed the enzyme immunoassay (EIA) kit which is a stable and convenient assay system for rat leptin in its plasma, serum and culture supernatant.
Quantitative determination kits for Biomarker Flyer download [PDF]
This ELISA kit is used for quantitative determination of rat leptin in its plasma, serum & culture supernatant samples. It has a lot of advantage to perform the assay, such as good quantification, no influence with other body fluid factors or physiological active substances and needlessness of sample pretreatment.
The EIA kit has high specificity to rat leptin and shows less than 0.02% 0.04% cross reactivity to human Leptin. And it has no cross reactivity with rat IL-1α, IL-1β, rat TNF-α, human TNF-α and other cytokines.
This EIA kit for determination of rat leptin in plasma, serum and culture supernatant samples is based on a sandwich enzyme immunoassay. The 96 wells plate is coated with anti rat leptin monoclonal antibody. Rat leptin standard or samples and HRP-labeled anti rat leptin polyclonal antibody are added to the wells for one step sandwich immunoreaction. During this immunoreaction, monoclonal antibody antigen HRP labeled antibody complex are formed. After rinsing out excess HRP labeled antibody, HRP enzyme activity is determined and the concentration of rat leptin is calculated.
|1. Antibody coated plate||MTP*1||1plate
|Anti rat Leptin monoclonal antibody|
|2. Rat Leptin standard||lyophilized||1 vial||Rat Leptin (20ng)|
|3. HRP labeled antibody||liquid||1 bottle
|HRP labeled antibody from rabbit|
|4. Substrate buffer||liquid||1 bottle
|0.015% Hydrogen Peroxide|
|5. OPD tablet||tablet||2 tablets||o-Phenylenediamine hydrochloride|
|6. Stopping solution||liquid||1 bottle
|7. Buffer solution A||liquid||1 bottle
|Phosphate buffer including animal serum|
|8. Buffer solution B||liquid||1 bottle
|Phosphate buffer including surfactant|
|9. Washing solution
|10.Adhesive foil||2 sheets|
MTP*1 …. . Microtiration plate