Brain-derived neurotrophic factor （BDNF） is a member of the NGF family of neurotrophic factors （also named neurotrophins） that are required for the differentiation and survival of specific neuronal subpopulations in both the central as well as the peripheral nervous system. The neurotrophin family is comprised of at least four proteins including NGF，BDNF，NT-3，and NT-4/5. These secreted cytokines are synthesized as prepropeptides that are proteolytically processed to generate the mature proteins. All neurotrophins have six conserved cysteine residues that are involved in the formation of three disulfide bonds and all share approximately 55% sequence identity at the amino acid level. Similarly to NGF，bioactive BDNF is predicted to be a non-covalently linked homodimer.Sample Type：Cell culture supernatants，serum，plasma，and tissueIntended Use：The Human BDNF BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human BDNF concentrations in cell culture supernatants，serum，plasma，and tissue.Sensitivity：7pg/mlRange：1000-15.625pg/mlSpecificity：Recognizes human BDNF.Sample Volume：100ul/wellTest Principle：This assay employs the Sandwich immunoassay technique. An anti-h BDNF monoclonal coating antibody is adsorbed onto microwells. BDNF 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 BDNF antibody is added and binds to BDNF captured by the first antibody. Following incubation unbound Biotinylated anti-h BDNF antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h BDNF antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of BDNF 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 BDNF standard dilutions and BDNF 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.
