Nesfatin Enzyme Immunoassay (EIA) BioAssay Kit is an in vitro quantitative assay for detecting Nesfatin peptide based on the principle of Competitive Enzyme Immunoassay. The microplate in the kit is pre-coated with anti-rabbit secondary antibody. After a blocking step and incubation of the plate with anti- Nesfatin antibody, both biotinylated Nesfatin peptide and peptide standard or targeted peptide in samples interacts competitively with the Nesfatin antibody. Uncompeted (bound) biotinylated Nesfatin peptide then interacts with Streptavidin-horseradish peroxidase (SAHRP), which catalyzes a color development reaction. The intensity of colorimetric signal is directly proportional to the amount of biotinylated peptide-SA-HRP complex and inversely proportional to the amount of Nesfatin peptide in the standard or samples. This is due to the competitive binding to Nesfatin antibody between biotinylated Nesfatin peptide and peptides in standard or samples. A standard curve of known concentration of Nesfatin peptide can be established and the concentration of Nesfatin peptide in the samples can be calculated accordingly. EIA-NES-1 detects 420aa Nesfatin protein. No other active isoforms have been reported. The minimum detectable concentration of Nesfatin is 147pg/ml or 3.09pM. Sample Type:Serum, plasma and cell culture media etc.Intended Use:The BioAssay kit is used for quantitative protein detection, establishing normal range and validation of antibody array results.Specificity:The capture antibody provided in this kit recognizes human, mouse and rat Nesfatin.Sensitivity:The intra-assay reproducibility is CV<10% and inter-assay is CV<12%. Range:The detection range for the kit is 0.1-1000ng/mlKit Components:Nesfatin Microplate (Item A), 1x96wellsWash Buffer Concentrate (Item B), 1x25mlStandard Nesfatin peptide (Item C), 2x vialsAssay Diluent A (Item D), 1x30mlAssay Diluent B (Item E), 1x15mlBiotinylated Nesfatin peptide (Item F), 2x vialsHRP-Streptavidin concentrate (Item G), 1x600ulTMB One-Step Substrate Reagent (Item H), 1x12mlStop Solution (Item I), 1x8mlPositive control (Item M), 1x vialAnti- Nesfatin polyclonal antibody (Item N), 2x vialsStorage 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.