Nerve growth factor (NGF) is a neurotrophic factor and neuropeptide primarily involved in the regulation of growth, maintenance, proliferation, and survival of certain target neurons. As reported, when the pheochromocytoma cell line PC12 is exposed to nerve growth factor (NGF), the cells respond over a period of a week by ceasing cell division and extending neurites (Greene and Tischler, 1976). The cells were grown in Ham's F12K containing 5% fetal calf serum and 10% horse serum on ?polylysine or collagen coated plates. When cells reached log phase growth , fresh medium was added together with 10ng/mL of NGF, then cells were observed by inverted microscope everyday.
Cell division ceasing and differentiation of PC12 cells after incubation with NGF (10ng/mL) for 6 days was shown in Figure1.Control group which received no NGF displayed no neurite outgrowth and cells multiply rapidly.

Nerve Growth Factor?(NGF) is the archetypal?neurotrophin?of a family of?polypeptides. It has long occupied a critical role in developmental and adult neurobiology for its many important regulatory functions on the survival, growth and differentiation of nerve cells in the peripheral and central nervous system. A functional binding ELISA assay was conducted to detect the interaction of recombinant human NGF and recombinant human CASP3. Briefly, NGF was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to CASP3-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-NGF pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 μL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human NGF and recombinant human CASP3 was shown in Figure 1, the EC50?for this effect is 0.08 ug/mL.