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Recombinant human NT-3 protein

QK058

Brand: Qkine

Neurotrophin 3 (NT-3) protein is part of the neurotrophin family and plays a crucial role in embryonic development and the maintenance and neuroprotection of the adult nervous system. NT-3 protein is used in cell culture to promote the differentiation and survival of specific neural subpopulations in both the central nervous system and peripheral nervous system such as sensory neurons, cortical neurons, and oligodendrocytes. It is also involved in the maintenance of endothelial cells and myocardial cells.

Qkine NT-3 protein is a non-covalently linked homodimer with a molecular weight of 27.3 kDa, animal origin-free (AOF), carrier-free, tag-free, and non-glycosylated to ensure its purity with exceptional lot-to-lot consistency. It is suitable for the culture of reproducible and high-quality cortical neurons and oligodendrocytes.

Qkine 3-for-2 product campaign

Currency: 

Product name Catalog number Pack size Price Price (USD) Price (GBP) Price (EUR)
Recombinant human NT-3 protein, 25 µg QK058-0025 25 µg (select above) $ 355.00 £ 225.00 € 298.00
Recombinant human NT-3 protein, 50 µg QK058-0050 50 µg (select above) $ 515.00 £ 380.00 € 444.00
Recombinant human NT-3 protein, 100 µg QK058-0100 100 µg (select above) $ 760.00 £ 560.00 € 655.00
Recombinant human NT-3 protein, 500 µg QK058-0500 500 µg (select above) $ 3,100.00 £ 2,300.00 € 2,687.00
Recombinant human NT-3 protein, 1000 µg QK058-1000 1000 µg (select above) $ 4,950.00 £ 3,600.00 € 4,205.00

Note: prices shown do not include shipping and handling charges.

Qkine company name and logo are the property of Qkine Ltd. UK.

Alternative protein names
NTF3, HDNF, NGF-2, NGF2, NT 3, NT3, neurotrophin 3, Nerve growth factor 2, Neurotrophic factor
Species reactivity

human

species similarity:
mouse – 100%
rat – 100%
porcine – 100%
bovine – 98%

Frequently used together with:

Summary

  • High purity human NT-3 protein (Uniprot: P20783)
  • >98%, by SDS-PAGE quantitative densitometry
  • 27.3 kDa dimer, 13.7 kDa monomer
  • Expressed in E. coli
  • Animal origin-free (AOF) and carrier protein-free
  • Manufactured in Cambridge, UK
  • Lyophilized from acetonitrile/TFA
  • Resuspend in 10 mM HCl (Reconstitution solution A) at >100 µg/ml, prepare single-use aliquots, add carrier protein if desired and store frozen at -20°C (short-term) or -80°C (long-term)
Handling and Storage FAQ

Featured applications

  • Neural differentiation
  • Neuronal maturation
  • Peripheral neuron differentiation
  • Oligodendrocyte differentiation
  • iPSC-derived dorsal root ganglion organoids generation
  • Human cortical organoid maturation
  • iPSC-derived sensory neuron differentiation

Bioactivity

Bioactivity graph showing the EC50 of 23 ng/ml (0.84 nM) for Qkine recombinant human NT-3

NT-3 protein bioactivity is measured using a luciferase reporter assay in HEK293T cells co-transfected with the TrkA receptor. Cells are treated in triplicate with a serial dilution of Qk058 for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. EC50 = 0.97 nM (26.481 ng/mL). Data from Qk058 lot #104407.

 

Purity

SDS-PAGE gel showing high purity reduced and non-reduced forms of NT-3

NT-3 protein migrates mainly as a single band at 13.7 kDa in non-reducing (NR) conditions and upon reduction (R). Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptoethanol, R) and non-reduced (-β-mercaptoethanol, NR) conditions and stained with Coomassie Brilliant Blue R-250. A faint band visible at 27.3 kDa in NR and R conditions, corresponds to the non-covalently linked NT-3 dimer. No contaminating bands are visible. Data from Qk058 batch #104407.

Further quality assays

  • Mass spectrometry: single species with expected mass
  • Endotoxin: <0.005 EU/μg protein (below level of detection)
  • Recovery from stock vial:  >95%

Qkine NT-3 is as biologically as a comparable alternative supplier protein

Quantitative luciferase assay with Qkine NT-3 (Qk058, green) and alternative supplier NT-3 (Supplier B, black). Cells were treated in triplicate with a serial dilution of NT-3 for 3 hours. Firefly luciferase activity was measured and normalized to control Renilla luciferase activity.

Protein background

Neurotrophin 3 protein is a member of the neurotrophic factor or neurotrophin family which includes nerve growth factor (NGF), brain–derived neurotrophic factor (BDNF) and neurotrophin–4/5 (NT–4/5) [1-3].

NT3 protein plays a crucial role in the development and maintenance of the nervous system. During embryonic development, it is involved in the growth and survival of sensory neurons, as well as in the formation of connections between neurons [4]. In the adult nervous system, it has a neuroprotective role where it is involved in the maintenance and repair of sensory neurons [5]. It supports their long-term survival and function. It promotes the survival and differentiation of peripheral, sensory, and spinal motor neurons, and oligodendrocytes [6-11]. NT3 protein is also involved the heart development and myocardial vasculature [12,13].

NT3 protein is used in cell culture to promote the differentiation and survival of specific neural subpopulations in both the central nervous system and peripheral nervous system [2]. It is used to differentiate induced pluripotent stem cells into cortical neurons such as GABAergic and glutamatergic neurons along with BDNF, glial-derived neurotrophic factor (GDNF), insulin-like growth factor 1 (IGF-1) [14-17]. Cortical organoids also require NT-3 for maturation [18].

In addition, it is a main growth factor for the differentiation of oligodendrocytes along with platelet-derived growth factor (PDGF-AA) and IGF-1 [19,20]. Finally, it can also be used to maintain endothelial cells and myocardial cells [12,13] .

NT-3 is part of the cysteine knot family of growth factors that share high structural homology with other neurotrophins such as NGF and BDNF. It is structurally characterized by the presence of six conserved cysteine residues that result in each protomer forming a twisted four-stranded beta-sheet, with three intertwined disulfide bonds. Bioactive NT3 is a non-covalently linked 27.3 kDa homodimer of two 13.6 kDa monomers [21].

Human NT3 protein cDNA encodes a 257 amino acid residue precursor protein with a signal peptide and a proprotein that is proteolytically processed to generate the 119 amino acid residue mature NT-3. The amino acid sequence of mature human NT-3 is identical to mice, rats, and pigs. It exerts its effects by binding to two different classes of transmembrane receptors on the surface of neurons. The primary receptor for NT-3 is TrkC (tyrosine receptor kinase C), although it can also interact with the low-affinity p75 neurotrophin receptor. In certain cells, it is also known to activate TrkA and TrkB kinase receptors [2].

Neurotrophin 3 and other neurotrophins such as NGF and BDNF have been studied for their potential therapeutic applications in treating various neurodegenerative diseases, spinal cord injuries, and peripheral nerve injuries [22-24].

Research is ongoing on the potent ability of NT3 protein to protect degenerating neurons and promote regeneration. This protein may also be a potential therapeutic target for depression and anxiety disorders by exerting its effect on neurotransmitters and the hypothalamic-pituitary-adrenal axis [24].

Background references

  1. Bates, B. et al. Neurotrophin–3 is required for proper cerebellar development. Nat. Neurosci. 2, 115–117 (1999). doi: 10.1038/5669
  2. Bibel, M. & Barde, Y.-A. Neurotrophins: key regulators of cell fate and cell shape in the vertebrate nervous system. Genes Dev. 14, 2919–2937 (2000). doi: 10.1101/gad.841400
  3. Skaper, S. D. The Neurotrophin Family of Neurotrophic Factors: An Overview. in Neurotrophic Factors: Methods and Protocols (ed. Skaper, S. D.) 1–12 (Humana Press, 2012). doi:10.1007/978-1-61779-536-7_1
  4. Maisonpierre, P. C. et al. NT-3, BDNF, and NGF in the developing rat nervous system: Parallel as well as reciprocal patterns of expression. Neuron 5, 501–509 (1990). doi.org/10.1016/0896-6273(90)90089-X
  5. Mattson, M. P. & Scheff, S. W. Endogenous Neuroprotection Factors and Traumatic Brain Injury: Mechanisms of Action and Implications for Therapy. J. Neurotrauma 11, 3–33 (1994). doi: 10.1089/neu.1994.11.3
  6. Goto, K. et al. Simple Derivation of Spinal Motor Neurons from ESCs/iPSCs Using Sendai Virus Vectors. Mol. Ther. Methods Clin. Dev. 4, 115 (2017). doi: 10.1016/j.omtm.2016.12.007
  7. Schwartzentruber, J. et al. Molecular and functional variation in iPSC-derived sensory neurons. Nat. Genet. 50, 54–61 (2018). doi: 10.1038/s41588-017-0005-8
  8. Marton, R. M. et al. Differentiation and maturation of oligodendrocytes in human three-dimensional neural cultures. Nat. Neurosci. 22, 484–491 (2019). doi: 10.1038/s41593-018-0316-9
  9. Stebbins, M. et al. Human pluripotent stem cell–derived brain pericyte–like cells induce blood-brain barrier properties. Sci. Adv. 5, eaau7375 (2019). doi: 10.1126/sciadv.aau7375
  10. Kamata, Y. et al. A robust culture system to generate neural progenitors with gliogenic competence from clinically relevant induced pluripotent stem cells for treatment of spinal cord injury. Stem Cells Transl. Med. 10, 398 (2021). doi: 10.1002/sctm.20-0269
  11. Yun, W. et al. OCT4-induced oligodendrocyte progenitor cells promote remyelination and ameliorate disease. Npj Regen. Med. 7, 1–15 (2022). doi: 10.1038/s41536-021-00199-z
  12. Caporali, A. & Emanueli, C. Cardiovascular Actions of Neurotrophins. Physiol. Rev. 89, 279 (2009). doi: 10.1152/physrev.00007.2008
  13. Delgado, A. C. et al. Endothelial NT-3 Delivered by Vasculature and CSF Promotes Quiescence of Subependymal Neural Stem Cells through Nitric Oxide Induction. Neuron 83, 572–585 (2014). doi: 10.1016/j.neuron.2014.06.015
  14. Vicario-Abejón, C., Owens, D., McKay, R. & Segal, M. Role of neurotrophins in central synapse formation and stabilization. Nat. Rev. Neurosci. 3, 965–974 (2002). doi: 10.1038/nrn988
  15. Zeng, H. et al. Specification of Region-Specific Neurons Including Forebrain Glutamatergic Neurons from Human Induced Pluripotent Stem Cells. PLOS ONE 5, e11853 (2010). doi: 10.1371/journal.pone.0011853
  16. Lin, L., Yuan, J., Sander, B. & Golas, M. M. In Vitro Differentiation of Human Neural Progenitor Cells Into Striatal GABAergic Neurons. Stem Cells Transl. Med. 4, 775–788 (2015). doi: 10.5966/sctm.2014-0083
  17. Fernandopulle, M. S. et al. Transcription-factor mediated differentiation of human iPSCs into neurons. Curr. Protoc. Cell Biol. 79, e51 (2018). doi: 10.1002/cpcb.51
  18. Gordon, A. et al. Long-term maturation of human cortical organoids matches key early postnatal transitions. Nat. Neurosci. 24, 331–342 (2021). doi: 10.1038/s41593-021-00802-y
  19. Hu, B.-Y., Du, Z.-W., Li, X.-J., Ayala, M. & Zhang, S.-C. Human oligodendrocytes from embryonic stem cells: conserved SHH signaling networks and divergent FGF effects. Dev. Camb. Engl. 136, 1443–1452 (2009). doi: 10.1242/dev.029447
  20. Wang, S. et al. Human iPSC-derived oligodendrocyte progenitors can myelinate and rescue a mouse model of congenital hypomyelination. Cell Stem Cell 12, 252–264 (2013). doi: 10.1016/j.stem.2012.12.002
  21. Butte, M. J., Hwang, P. K., Mobley, W. C. & Fletterick, R. J. Crystal Structure of Neurotrophin-3 Homodimer Shows Distinct Regions Are Used To Bind Its Receptors,. Biochemistry 37, 16846–16852 (1998). doi: 10.1021/bi981254o
  22. Rangasamy, S. B., Soderstrom, K., Bakay, R. A. E. & Kordower, J. H. Chapter 13 – Neurotrophic factor therapy for Parkinson’s disease. in Progress in Brain Research (eds. Björklund, A. & Cenci, M. A.) vol. 184 237–264 (Elsevier, 2010). doi: 10.1016/S0079-6123(10)84013-0
  23. Keefe, K. M., Sheikh, I. S. & Smith, G. M. Targeting Neurotrophins to Specific Populations of Neurons: NGF, BDNF, and NT-3 and Their Relevance for Treatment of Spinal Cord Injury. Int. J. Mol. Sci. 18, 548 (2017). doi: 10.3390/ijms18030548
  24. de Miranda, A. S., de Barros, J. L. V. M. & Teixeira, A. L. Is neurotrophin-3 (NT-3): a potential therapeutic target for depression and anxiety? Expert Opin. Ther. Targets 24, 1225–1238 (2020). doi: 10.1080/14728222.2020.1846720