Leptin is a product of the mouse obese gene. Mice with mutations in the obese gene that block the synthesis of leptin are obese and diabetic，and have reduced activity，metabolism，and body temperature. Rat leptin shares approximately 96% and 82% sequence identity with the mouse and human protein，respectively. The expression of leptin mRNA is restricted to adipose tissue.Sample Type：Cell culture supernatants，serum，plasma，and tissueIntended Use：The Human Leptin BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human Leptin concentrations in cell culture supernatants，serum，plasma，and tissue.Sensitivity：60pg/mlRange：125-4000pg/mlSpecificity：Recognizes human Leptin.Sample Volume：100ul/wellTest Principle：This assay employs the Sandwich immunoassay technique. An anti-h Leptin monoclonal coating antibody is adsorbed onto microwells. Leptin present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h Leptin antibody is added and binds to Leptin captured by the first antibody. Following incubation unbound Biotinylated anti-h Leptin antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h Leptin antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of Leptin present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven Leptin standard dilutions and Leptin sample concentration determined.Kit Components：Aluminium pouches with a Microwell Plate： 8x12Standard： 2xVialBiotinylated antibody： 2xVialStreptavidin-HRP： 2xVialAssay Buffer （25ml/Vial）： 2xVialWash Buffer Concentrate 20x （30ml/Vial）： 1xVialTMB Substrate Solution： 1x12mlStop Solution （12ml/Vial）： 1xVialPlate Covers- Adhesive strips： 3xstripsInstruction： 1xStorage and Stability：Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product，centrifuge the original vial after thawing and prior to removing the cap.
