Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Neurotrophin-4 (hNT-4) #5592

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Source

Recombinant human Neurotrophin-4 (hNT-4) Gly81-Ala210 (Accession #NP_006170) was produced in E.coli at Cell Signaling Technology.

Molecular Characterization

Recombinant hNT-4 does not have a Met on the amino terminus and has a calculated MW of 13,923. DTT-reduced and non-reduced protein migrate as 14 kDa polypeptides. The expected amino-terminal GVSET of recombinant hNT-4 was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hNT-4. All lots are greater than 98% pure.

Bioactivity

The activity of hNT-4 was assessed by quatification of phospho-TrkB in TrkB transfected NIH/3T3 cells. The observed ED50 values are between 1-4 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hNT-4 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hNT-4 and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The activity of hNT-4 was assessed. After starvation, TrkB transfected NIH/3T3 cells were treated with increasing concentrations of hNT-4 for 5 minutes. Cells were lysed, and phospho-TrkB was quatified using PathScan® Phospho-TrkB (panTyr) Sandwhich ELISA Kit #7108. OD450 is shown.

Western Blotting

Western Blotting

Western blot analysis of extracts from TrkB transfected NIH/3T3 cells untreated and treated with increasing concentrations of hNT-4 for 5 minutes, using Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb #4621 (upper) or TrkB (80E3) Rabbit mAb #4603 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hNT-4.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl and 20 μg BSA per 1 μg hNT-4. Carrier free: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl.

Background

NT-4 is a member of the structurally related neurotrophin family of proteins, which includes β-NGF, BDNF and NT-3 (1). NT-4 is expressed in a number of cell types and tissues, including neuronal cells, normal breast epithelial cells, melanocytes, activated T cells, and granulocytes (1-5). NT-4 is required for the development of peripheral sensory neurons (6,7). NT-4 may be important for the development of long term memory (8). Increased NT-4 expression in melanoma cells promotes cell proliferation and migration (5). NT-4 is secreted from cells as a precursor protein, which is proteolytically cleaved into the mature form (1). NT-4 signaling is mediated through two distinct receptors, the neurotrophin receptor p75NTR and the Trk tyrosine kinase receptor TrkB. While all neurotrophins bind to the p75NTR receptor, NT-4 preferentially binds to the TrkB receptor (1).

  1. Ip, N.Y. et al. (1992) Proc Natl Acad Sci U S A 89, 3060-4.
  2. Vanhecke, E. et al. (2011) Clin Cancer Res 17, 1741-52.
  3. Moalem, G. et al. (2000) J Autoimmun 15, 331-45.
  4. Laurenzi, M.A. et al. (1998) J Leukoc Biol 64, 228-34.
  5. Truzzi, F. et al. (2008) J Invest Dermatol 128, 2031-40.
  6. Conover, J.C. et al. (1995) Nature 375, 235-8.
  7. Liu, X. et al. (1995) Nature 375, 238-41.
  8. Xie, C.W. et al. (2000) Proc Natl Acad Sci U S A 97, 8116-21.

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