Nesfatin-1 is a naturally occurring protein molecule produced by the brains of mammals. It is responsible for regulating appetite and production of body fat. Nesfatin-1 is a metabolic polypeptide encoded in the N-terminal region of the protein precursor, Nucleobindin2(NUCB2). Recombinant human Nesfatin-1 is a 9.7kD protein containing 82aa residues. Originally identified as a hypothalamic neuropeptide, Nesfatin-1 is also expressed in other areas of the brain, and in pancreatic islets of Langerhans, gastric endocrine cells and adipocytes. Nesfatin-1 plays a role in hunger and energy regulation in an independent manner.
Sample Type:Cell culture supernates and serum.
Intended Use:For quantitative detection of human Nesfatin-1 in cell culture supernates and serum.
Sensitivity:<10pg/ml
Range:31.2-2000pg/ml
Specificity:Natural and recombinant mouse CCL8
Test Principle:This human Nesfatin-1 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Nesfatin-1 has been precoated onto 96-well plates. Standards(E.coli) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Nesfatin-1 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 Nesfatin-1 amount of sample captured in plate.
Kit Components:96-well plate precoated with human Nesfatin-1 antibody, 1Lyophilized recombinant human Nesfatin-1 standard, 2ng/tube×2Biotinylated human Nesfatin-1 antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB stop solution, 10ml
Storage 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.
