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.

14.2

In culture medium, buffer or water. NGF (2.5S) is soluble at ≥1mg/ml in neutral and acidic solutions.

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.

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.

Male mouse submaxillary glands.

~98%. Purified by the method of Bocchini and Angeletti.

~1mg/ml (after reconstitution)

Supplied as a sterile filtered lyophilized powder. Reconstitute with sterile culture medium, buffer or water. NGF (2.5S) is soluble at > 1mg/ml in neutral and acidic solutions

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.