Leptin, Human (Ob, Obese Protein, Obesity Factor) BioAssay(TM) ELISA Kit

This Human Leptin ELISA kit is to be used for the in vitro quantitative determination of leptin in human serum, plasma, buffered solution or cell culture medium. The assay recognizes both natural and recombinant forms of leptin. Leptin, the product of the ob gene, is a hormone secreted by adipocytes. Animals with mutations in the ob gene are obese, diabetic and have reduced activity. Administration of recombinant leptin to these animals decreases food intake and causes weight loss. In humans, this type of mutation has not been found. Human leptin cDNA encodes a 167 amino acid non-glycosylated protein including a 21aa signal peptide which is cleaved to give mature human leptin. The human receptor for leptin (OB- B-R) has been identified as a 1144 amino acid transmembrane glycoprotein. t is expressed in the choroid plexus and in the hypothalamus. Leptin is implicated in an increasing number of endocrine regulations including adiposity, satiety, energy homeostasis, puberty and fertility. Little is known about the physiologic actions of leptin in humans. The availability of a sensitive and specific assay to measure concentrations of leptin in serum and plasma will accelerate our understanding. Principle: This Human Leptin kit is a solid phase sandwich ELISA. A monoclonal antibody specific for human leptin has been coated onto the wells of the microtiter strips provided. Samples, including standards of known leptin content, control specimens, and unknowns, are pipetted into these wells followed by addition of a second biotinylated monoclonal antibody. During the first incubation, the leptin antigen binds to the immobilized (capture) antibody on one site and to the solution phase biotinylated antibody on a second site. After removal of excess secondary antibody, Streptavidin-Peroxidase enzyme is added. This binds to the biotinylated antibody to complete the four-member sandwich. After a second incubation and washing to remove all the unbound enzyme, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of leptin present in the original specimen. Sensitivity: <3.5pg/ml , Range: , 15.6-1000pg/ml, Incubation Time: 3 hours at RT, Sample size: 100ul, Accuracy: Inter-assay CV: <5.3%, Intra-assay CV: <3.9%, Kit Components: 96 Test Kit 192 Test Kit, L1670-14A Standard 2 vials 4 vials, L1670-14B Standard Diluent 1x60ml 2x60ml, L1670-14C Microtiter Plate 1x96wells 2x96wells, L1670-14D Mab (Biotin) 1x11ml 2x11ml, L1670-14E Streptavidin (HRP), 100X 1x125ul 2x125ul , L1670-14F HRP Diluent 1x25ml 1x25ml, L1670-14G Wash Buffer, 25X 1x100ml 1x100ml, L1670-14H TMB 1x25ml 1x25ml, L1670-14J Stop Solution 1x25ml 1x25ml, Storage and Stability: Store all components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.