Active Nerve Growth Factor (NGF)

NGFB; Beta-NGF; HSAN5; NGF-B; Beta-Nerve Growth Factor

ACTIVITY TEST

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.

NGF is also involved in pathways besides those regulating the life cycle of neurons. Besides, Alpha-2-Macroglobulin (a2M) has been identified as an interactor of NGF, thus a binding ELISA assay was conducted to detect the interaction of recombinant human NGF and recombinant human a2M. Briefly, NGF were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to a2M-coated microtiter wells and incubated for 2h 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, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of NGF and a2M was shown in Figure 2, and this effect was in a dose dependent manner.
Figure. The binding activity of NGF with a2M

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
The Tohoku Journal of Experimental Medicine Low-intensity aerobic exercise mitigates exercise-induced bronchoconstriction by improving the function of adrenal medullary chromaffin cells in asthmatic rats Pubmed:25212248
International Journal of Pharmaceutics Neurotrophic and antioxidant effects of silymarin comparable to 4-methylcatechol in protection against gentamicin-induced ototoxicity in guinea pigs Pubmed:25445537
Yonsei Medical Journal NGF and HB-EGF: Potential Biomarkers that Reflect the Effects of Fesoterodine in Patients with Overactive Bladder Syndrome Pubmed:25510766
Molecular and Cellular Endocrinology High glucose upregulates CYP24A1 expression which attenuates the ability of 1,25(OH)2D3 to increase NGF secretion in a rat Schwann cell line RSC96 Pubmed:25614971
Mol Cell Endocrinol High glucose upregulates CYP24A1 expression which attenuates the ability of 1,25(OH)2D3 PubMed: 25614971
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Neurotoxicity Research Pro-apoptotic Action of Corticosterone in Hippocampal Organotypic Cultures Pubmed:27189478
Psychoneuroendocrinology Chronic mild stress influences nerve growth factor through a matrix metalloproteinase-dependent mechanism Pubmed:26771945
Genet Mol Res Changes in growth factor levels in the cerebrospinal fluid of autism patients after transplantation of human umbilical cord blood mononuclear cells and umbilical cord-derived mesenchymal stem cells Pubmed:27323064
The Journal of Maternal-Fetal & Neonatal Medicine Differential expression of cord blood neurotrophins in gestational diabetes: the impact of fetal growth abnormalities pubmed:28081639
IOS Press Content Library An Intranasal Formulation of Erythropoietin (Neuro-EPO) Prevents Memory Deficits and Amyloid Toxicity in the APPSwe Transgenic Mouse Model of Alzheimer's … articles:journal-of-alzheimers-disease
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Progress in Neuro-Psychopharmacology & Biological Psychiatry Antidepressant drugs for beta amyloid-induced depression: A new standpoint? pubmed:28499898
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Experimental Neurology Preterm umbilical cord blood derived mesenchymal stem/stromal cells protect preterm white matter brain development against hypoxia-ischemia Pubmed:30012511
Biochemical Pharmacology Effects of n-3 PUFA enriched and n-3 PUFA deficient diets in naïve and Aβ-treated female rats Pubmed:30028991
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