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2.5S NERVE GROWTH FACTOR (NGF) (non-HPLC purified) CLMCNET-005.25 Lot : SP0507 DESCRIPTION: Cedarlane® purified NGF is prepared from male mouse submandibular gland, and is useful for the study of neuronal growth and differentiation in culture and in vivo. In the peripheral nervous system, Nerve Growth Factor (NGF) promotes the survival and growth of sympathetic and spinal sensory neurons1. In the central nervous system, NGF enhances survival and differentiation of cholinergic neurons particularly in the basal forebrain2. NGF also appears to be a mitotic agent for mast cells3 and for basophilic leukocytes4. These preparations of NGF have been used to study the development of sympathetic neurons7, and to examine proliferation of mast cells3 and leukocytes4. PRESENTATION: 250 µg, lyophilized, sterile, and sealed in glass vials under vacuum. RECONSTITUTION: The 2.5S NGF is lyophilized from an acetic acid solution and should be reconstituted in dilute acetic acid. Bring into solution by injecting 10 – 50 µl of 0.02% acetic acid into the vial. Then add working buffer and aliquot. STORAGE AND STABILITY: Lyophilized form is stable at 4°C for a short term. Lyophilized and reconstituted format is stable at -20°C. Prepare aliquots appropriate for single use to avoid repeated freezing and thawing. PURITY: Current protocols for the purification of NGF are described in detail by Darling and Shooter5. This 2.5S NGF is prepared from male mouse submandibular glands by the method of Mobley et al6. This method produces several NGF chains reflecting the lack of the NH2-terminal octapeptide and/or the COOH-terminal arginine residues. Purity of this product is >90%. ACTIVITY: The ED50 of this lot of 2.5S NGF (Lot# SP0507) is 0.17 ng/ml as determined by bioassay using dissociated mouse sympathetic neurons7. Maximal activity in the bioassay is reached at approximately 1 to 3ng/ml 2.5S NGF. Continued Overleaf.......... REFERENCES: 1. 2. 3. 4. 5. 6. 7. Levi-Montalcini, R. (1987). The nerve growth factor 35 years later. Science 237:1154-1157. Hefti, F. and Weiner, W.J. (1986). Nerve growth factor and Alzheimer’s disease. Ann.Neurol. 20:275281. Aloc, L. and Levi-Montalcini, R. (1977). Mast cells increase in tissues of neonatal rats injected with nerve growth factor. Brain Res. 133:358-366. Bienenstock, J., Tomioka, M., Matsuda, H., Stead, R.H., Quinonez, G., Simon, G.T., Coughlin, M.D., and Denburg, J.A. (1987). The role of mast cells in inflammatory processes: Evidence for nerve/mast cell interactions. Int.Arch.Allergy Appl.Immunol. 82:238-243. Matsuda, H., Coughlin, M.D., Bienenstock, J., and Denburg, J.A. (1988). Nerve growth factor promotes human hemopoietic colony growth and differentiation. Proc.Natl.Acad.Sci. USA, 85:6508-6512. Darling, T.L. and Shooter, E.M. (1984). In Cell Culture Methods for Molecular and Cellular Biology (D.W. Barnes, D.A. Sirbasku, and G.H. Sato, eds.), vol. 4, Liss, New York, pp 79-93. Mobley, W.C., Schenker, A., and Shooter, E.M. (1976) Characterization and isolation of proteolytically modified nerve growth factor. Biochemistry, 15, 5543-51. Coughlin, M.D. and Collins, M.B., (1985) Nerve growth factor-independent development of embryonic mouse sympathetic neurons in dissociated cell culture. Dev. Biol. 110, 392-401. Laboratory Reagent For Research Use Only JV 04/14/11