Revision 3

#5237Store at -20C

10 µg

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
MW (kDa):

14

UniProt ID:

#P20783

Entrez-Gene Id:

4908

Background

NT-3 is a member of the structurally related neurotrophin family of proteins, which includes β-NGF, BDNF and NT-4 (1). NT-3 is expressed in a number of cell types including neuronal cells, eosinophils, and melanocytes (1-3). NT-3 is required for the development of peripheral sensory neurons (4). NT-3 is secreted from cells as a precursor protein, which is proteolytically cleaved into the mature form (1). NT-3 signaling is mediated through two distinct receptors, the neurotrophin receptor p75NTR and the Trk tyrosine kinase receptor TrkC. While all neurotrophins bind to the p75NTR receptor, NT-3 preferentially binds to TrkC and, with lesser affinity, TrkB (1).

  1. Huang, E.J. and Reichardt, L.F. (2001) Annu Rev Neurosci 24, 677-736.
  2. Noga, O. et al. (2003) Clin Exp Allergy 33, 649-54.
  3. Truzzi, F. et al. (2008) J Invest Dermatol 128, 2031-40.
  4. Tessarollo, L. et al. (1994) Proc Natl Acad Sci U S A 91, 11844-8.

Endotoxin

Less than 0.01 ng endotoxin/1 μg hNT-3.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hNT-3. All lots are greater than 98% pure.

Source / Purification

Recombinant human Neurotrophin-3 (hNT-3) Tyr139-Thr257 (Accession #NP_002518) was produced in E.coli at Cell Signaling Technology.

Bioactivity

The bioactivity of recombinant hNT-3 was determined in a TrkC-transfected NIH/3T3 cell proliferation assay. The ED50 of each lot is between 0.5 - 5 ng/ml.

Background

NT-3 is a member of the structurally related neurotrophin family of proteins, which includes β-NGF, BDNF and NT-4 (1). NT-3 is expressed in a number of cell types including neuronal cells, eosinophils, and melanocytes (1-3). NT-3 is required for the development of peripheral sensory neurons (4). NT-3 is secreted from cells as a precursor protein, which is proteolytically cleaved into the mature form (1). NT-3 signaling is mediated through two distinct receptors, the neurotrophin receptor p75NTR and the Trk tyrosine kinase receptor TrkC. While all neurotrophins bind to the p75NTR receptor, NT-3 preferentially binds to TrkC and, with lesser affinity, TrkB (1).

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Revision 3
    #5237

    Human Neurotrophin-3 (hNT-3)

    Human Neurotrophin-3 (hNT-3): Image 1 Expand Image
    对 hNT-3 诱导 TrkB 磷酸化的能力进行评估。饥饿后,转染 TrkB 的 NIH/3T3 细胞用浓度递增的 hNT-3 处理 5 分钟。细胞裂解后,使用 PathScan® Phospho-TrkB (panTyr) Sandwhich ELISA Kit #7108 对磷酸-TrkB 进行定量分析。OD450-OD650 如图所示。
    Human Neurotrophin-3 (hNT-3): Image 2 Expand Image
    对用浓度递增的 hNT-3 处理且转染 TrkC 的 NIH/3T3 细胞的增殖进行评估。处理 24 小时后,细胞用 BrdU 标记 4 小时。通过 ELISA 确定 BrdU 的含量,以确定 OD450-OD690
    Human Neurotrophin-3 (hNT-3): Image 3 Expand Image
    对 6 µg 还原 (+) 和非还原 (-) 重组 hNT-3 进行 SDS-PAGE 并用考马斯蓝染色过夜来测定重组 hNT-3 的纯度。
    Human Neurotrophin-3 (hNT-3): Image 4 Expand Image
    使用 Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb #4621(上图)或 TrkB (80E3) Rabbit mAb #4603(下图),对未经处理和已经 hNT-3 处理 5 分钟且转染 TrkB 的 NIH/3T3 细胞的提取物进行蛋白质印迹分析。