Description
Principle of the assay: human Leptin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Leptin has been precoated onto 96-well plates. Standards(E.coli, V22-C167) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Leptin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Leptin amount of sample captured in plate.
Background: Leptin (or obese, OB) is a circulating hormone that is expressed abundantly and specifically in the adipose tissue.1 Human ob protein is a 166-amino acid polypeptide with a putative signal sequence and is 84 and 83% homologous to the mouse and rat ob proteins, respectively.2 Northern blot analysis reveals that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart.3 Indeed, Leptin is first described as an adipocyte-derived signalling factor, which, after interaction with its receptors, induces a complex response including control of bodyweight and energy expenditure. Leptin seems in addition to its role in metabolic control to have important roles in reproduction and neuroendocrine signalling.4 Leptin is a powerful inhibitor of bone formation in vivo.5 Comparative mapping of mouse and human DNA indicates that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q.3 The standard product used in this kit is recombinant human Leptin