Nerve growth factor(NGF) is a polypeptide involved in the regulation of growth and differentiation of sympathetic and certain sensory neurons. NGF is thought to have a profound effect on the development and maintenance of sympathetic and embryonic sensory neurones. NGF activity isolated from the male Monkey submaxillary gland(MSG) consists of three types of subunits, alpha, beta and gamma, which specifically interact to form a 7S, approximately 130,000-molecular weight(Mr) complex. The 7S complex contains two identical 118aa beta-chains, which are solely responsible for the nerve growth-stimulating activity of NGF. NGF, which is expressed by inflammatory cells and effects changes that lead to increased neural responsiveness, could be a pivotal mediator in allergic rhinitis. The standard product used in this kit is monkey 2.5S NGF, which is a dimmer linking with two polypeptide chains of 120aa. Sample Type: Cell culture supernates, serum. Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Monkey NGF/NGF beta. Sensitivity: <1pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Monkey NGF/NGF beta. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Monkey NGF/NGF Beta BioAssay(TM) ELISA Kit was based on standard sandwich enzyme- linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Monkey NGF/NGF Beta has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: S122-A241) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Monkey NGF/NGF Beta 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 Monkey NGF/NGF Beta amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Monkey NGF/NGF beta, 1x plate Lyophilized recombinant Monkey NGF/NGF beta standard, 2x10ng/tube Biotinylated anti-Monkey NGF/NGF beta, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
