Brain-derived neurotrophic factor (BDNF) is a prosurvival factor induced by cortical neurons that is necessary for survival of striatal neurons in the brain. It is a secreted protein with the molecular weight of 27.8kD, consisting of 247 amino acids. It is known to promote neuronal survival and differentiation. BDNF shares substantial amino acid sequence identity with nerve growth factor (NGF). BDNF and neurotrophin-3 (NT-3) are two recently cloned neurotrophic factors that are homologous to NGF. mRNA products of the BDNF and NT-3 genes are detected in the adult human brain, suggesting that these proteins are involved in the maintenance of the adult nervous system.1 BDNF and other neurotrophins are critically involved in long-term potentiation (LTP). BDNF-mediated LTP is induced postsynaptically. BDNF has trophic effects on serotonergic (5-HT) neurons in the central nervous system. BDNF has an essential maintenance function in the regulation of anxiety-related behavior and in food intake through central mediators in both the basal and fasted state. It plays a role in treating breathing disorders such as respiratory insufficiency after spinal injury. Intended Use: BDNF BioAssay(TM) ELISA Kit is a sandwich ELISA for quantitative detection of rat BDNF in cell culture supernates, cell lysates, serum and plasma (heparin, EDTA, citrate). Range: 31.2pg/ml-2000pg/ml Sensitivity: <15pg/ml Specificity: Natural and recombinant rat BDNF Crossreactivity: There is no detectable cross-reactivity with other relevant proteins. Kit Components: 143727A: Microtiter Strips, 1x96 wells 143727B: BDNF Standard, 2x10ng 143727C: Anti-rat BDNF (Biotin), 1x130ul (1:100 dilution) USB Cat No Kit Component 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml Storage and Stability: Store *143727B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add samples and standards and incubate the plate at 37C for 90 minutes. Do not wash. 2. Add biotinylated antibodies and incubate the plate at 37C for 60 minutes. Wash plate 3 times with 0.01M TBS or PBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5 times with 0.01M TBS or PBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 15-20 minutes. 5. Add TMB stop solution and read.
