Nerve Growth Factor, 2.5S, Murine (NGF)

Nerve growth factor has a variety of effects on the growth and development of sensory and sympathetic neurons. In the peripheral nervous system, NGF is required for the development and maintenance of sympathetic nerve cells that use catecholamine neurotransmitters. Molecular weight of the monomer is 13kD. Biological Activity: Assayed using the neurite outgrowth effect on PC-12 rat pheochromocytoma cells or chick embryo dorsal root ganglia. The minimum concentration needed for neurite outgrowth of PC-12 rat pheochromocytoma cells is 10ng/ml. Results may vary depending on cell line used. Extinction Coefficient (1%,280nm, pH7.0): 14.2 , Reconstitution: , In culture medium, buffer or water. NGF (2.5S) is soluble at >1mg/ml in neutral and acidic solutions. Storage and Stability: Lyophilized powder may be stored at -20°C. Reconstitute to nominal volume by adding sterile culture medium, buffer or water. Aliquot and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. NGF Assay Protocol: 1. Coat 6 well dishes with rat tail collagen Type I at a concentration of 2.5-5ug/cm2. Incubate 1 hour at 37°C. Aspirate and rinse 2 times with PBS before adding cell suspension. , 2. Prepare a cell suspension to yield 2x10e4 cells/ml in RPMI-1640/10% horse serum/ 5% FBS. Add 2ml of cell suspension to each well. , 3. Rehydrate NGF as described above to a final concentration of 1ug/ml. Add appropriate amounts of NGF to test at 0, 5, 10 and 20ng/ml final concentration. , 4. Incubate plates at 37°C for 3-5 days. , 5. Record neurite outgrowth at each NGF concentration.