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Qkine recombinant human NT3 protein vial
Bioactivity graph showing the EC50 of 23 ng/ml (0.84 nM) for Qkine recombinant human NT-3
SDS-PAGE gel showing high purity reduced and non-reduced forms of NT3 protein

Recombinant human/mouse/rat/porcine NT-3 protein (Qk058)

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.

Recombinant Neurotrophin 3 protein is a non-covalently linked homodimer with a molecular weight of 27.3 kDa.

Qkine NT-3 protein is 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.

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1000µg will be despatched as 2 x 500µg

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Summary

  • High purity human NT3 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 10mM 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)

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

NTF3, HDNF, NGF-2, NGF2, NT 3, NT3, neurotrophin 3, Nerve growth factor 2, Neurotrophic factor

Human
Mouse
Rat
Porcine (pig)

Frequently used together

Recombinant human BDNF protein (Qk050)
Recombinant human GDNF protein (Qk051)
Recombinant human PDGF-AA protein (Qk043)
Recombinant human/bovine/porcine IGF-1 protein (Qk047)
Recombinant human EGF protein (Qk011)
Recombinant human HGF (NK1) protein (Qk013)
Recombinant human VEGF 165 protein (Qk048)

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%

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine.  All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

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].

[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

Additional resources

FAQ

NT-3, or Neurotrophin-3, is a growth factor which belongs to the neurotrophin family of proteins.

NT3 protein plays a crucial role in the embryonic development and the maintenance of the nervous system. It supports the growth, survival, and differentiation of sensory neurons, cortical neurons, and oligodendrocytes.

The gene that encodes for Neurotrophin-3 is the NTF3 gene.

Neurotrophin proteins regulate neuronal growth and survival, neuroprotection, neurotransmitter release, synaptic function, and plasticity within the adult central and peripheral nervous system.

Neurotrophins promote the survival and growth of neurons by inhibiting apoptosis (programmed cell death) in a process called neuroprotection.

NT3 protein binds to specific receptors on the surface of neurons, primarly TrkC (tyrosine receptor kinase C), although it can also interact with the low-affinity p75 neurotrophin receptor.

Neurotrophin proteins include Nerve Growth Factor (NGF), Brain-Derived Neurotrophic Factor (BDNF), Neurotrophin-3 (NT-3), and Neurotrophin-4/5 (NT-4/5).

Our products are for research use only and not for diagnostic or therapeutic use.  Products are not for resale.

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