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Recombinant human GDNF protein

QK051

Brand: Qkine

Glial cell line-derived neurotrophic factor (GDNF) is a member of neurotrophin family and GDNF family of ligands (GFL). GDNF plays a crucial role in the development, growth, and survival of neurons in particular midbrain dopaminergic neurons. GNDF is used to maintain neurons and cortical organoids and to differentiate dopaminergic neurons from human pluripotent stem cell-derived neural progenitors. GDNF also facilitates the differentiation of neural progenitors to astrocytes.

Recombinant human GDNF bioactive 30 kDa homodimer. This protein is animal origin-free (AOF), carrier protein-free, and tag-free to ensure its purity with exceptional lot-to-lot consistency.

Currency: 

Product name Catalog number Pack size Price Price (USD) Price (GBP) Price (EUR)
Recombinant human GDNF protein, 25 µg QK051-0025 25 µg (select above) $ 280.00 £ 205.00 € 240.00
Recombinant human GDNF protein, 50 µg QK051-0050 50 µg (select above) $ 410.00 £ 305.00 € 357.00
Recombinant human GDNF protein, 100 µg QK051-0100 100 µg (select above) $ 620.00 £ 455.00 € 532.00
Recombinant human GDNF protein, 500 µg QK051-0500 500 µg (select above) $ 2,500.00 £ 1,840.00 € 2,150.00
Recombinant human GDNF protein, 1000 µg QK051-1000 1000 µg (select above) $ 3,950.00 £ 2,900.00 € 3,388.00

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
Astrocyte-derived trophic factor, ATF, ATF1, ATF2; glial cell line derived neurotrophic factor; glial derived neurotrophic factor; HFB1-GDNF; HSCR3
Species reactivity

human

species similarity:
mouse – 93%
rat – 93%
porcine – 94%
bovine – 92%

Frequently used together with:

Summary

  • High purity GDNF (Uniprot: P39905)
  • >98%, by SDS-PAGE quantitative densitometry
  • 15.1 kDa monomer, 30.4 kDa (dimer)
  • Expressed in E. coli
  • Animal origin-free (AOF) and carrier protein-free
  • Manufactured in Cambridge, UK
  • Lyophilized from acetonitrile, TFA
  • Resuspend in water at >100 µg/ml, prepare single use aliquots, add carrier protein if desired and store frozen at -20°C or -80°C
Handling and Storage FAQ

Featured applications

  • Differentiation of midbrain dopaminergic neurons

Bioactivity

Zebrafish FGF2 / bFGF Qk002 protein bioactivity lot #011

GDNF activity is determined using a SH-SY5Y cell proliferation assay. Cells were incubated with different concentrations of GDNF in the presence of retinoic acid and recombinant GFR α1 for 3 days before viable cell measurement using an MTS assay. Data are n=2. EC50 = 18 ng/ml. Data from Qk051 batch #104372.

Purity

Zebrafish FGF2 / bFGF Qk002 protein purity SDS-PAGE lot #011

GDNF migrates as a single band at 30 kDa in non-reducing (NR) conditions and 15 kDa upon reduction (R).  No contaminating protein bands are visible. Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk051 batch #104372.

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%

Comparison with other suppliers

Qkine GDNF (Qk051) bioactivity was compared with an alternative supplier (Supplier 1) using a SH-SY5Y cell proliferation assay (as above). Qkine GDNF was found to have higher bioactivity than Supplier 1 GDNF. Qk051 EC50 = 18 ng/ml, Supplier 1 EC50 = 26 ng/ml, P = 0.018

Protein background

Glial cell line-derived neurotrophic factor (GDNF) is a neurotrophic factor and a member of the GDNF family of ligands (GFL) which include neurturin, artemin, and persephin [1,2]. GFLs regulate several biological processes including cell survival, cell proliferation, cell differentiation, cell migration, and neurite outgrowth [1,3,4]. GDNF protein plays a crucial role in the development, growth, and survival of neurons in particular midbrain dopaminergic neurons [5]. It promotes the axon growth and innervation of dopamine neurons. It also facilitates the differentiation of neural progenitors to astrocytes and is involved in neuroprotection[6,7]. In addition, it is involved in kidney development and spermatogenesis [8,9].

In cell culture, GDNF supports the survival and growth of neurons and astrocytes. It is used in combination with other growth factors to differentiate human induced pluripotent stem cell-derived neural progenitors into neurons and astrocytes. GDNF is used for the differentiation, survival, and maturation of dopaminergic neurons with brain-derived neurotrophic factor (BDNF),FGF-8a/FGF-8b, and Shh [10-13]. Glutamatergic and GABAergic neurons are obtained with combinations of BDNF, GDNF, NT-3, or insulin-like growth factor 1 (IGF-1) [14-16]. Also, the culture of cortical organoids requires BDNF and GDNF for maturation [17,18]. Finally, it can be used to generate and maintain astrocytes along with ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF) [19-21].

GDNF is mainly produced and released by glial cells such as astrocytes, Schwann cells, and satellite cells [22]. It is a homodimer of a molecular weight of 30 kDa which belongs to the cystine-knot protein family [1,23]. The GFLs act as biologically active homodimers that signal through the transmembrane RET receptor tyrosine kinase [24]. GFL activation of RET is dependent upon co-receptors, namely the four GDNF Family Receptor α (GFRα1-4). GFL signalling specificity arises from the preferentially binding of each ligand to one of the four GFRα receptors; GDNF preferentially binds GFRα1 with high affinity [1,24].

GDNF has shown promise in various therapeutic applications neurodegenerative diseases and disorders such as Parkinson’s disease, amyotrophic lateral sclerosis, Huntington’s disease, peripheral nerve and spinal cord injuries [1,25]. The idea is to harness GDNF for clinical use to promote the survival and function of dopaminergic neurons, potentially slowing or reversing the progression of these diseases. Clinical trials using it has shown improved motor function in patients with Parkinson’s disease [26,27]. Finally, GDNF has also been proposed for the treatment of drug addiction and alcoholism [25].

Background references

  1. Airaksinen, M. S. & Saarma, M. The GDNF family: Signalling, biological functions and therapeutic value.Nat. Rev. Neurosci.3, 383–394 (2002).doi: 10.1038/nrn812
  2. von dem Bussche, M. & Tuszynski, M. H. Growth Factors: Neuronal Atrophy. inEncyclopedia of Neuroscience(ed. Squire, L. R.) 987–992 (Academic Press, 2009). doi:10.1016/B978-008045046-9.00143-1.
  3. Airaksinen, M. S., Titievsky, A. & Saarma, M. GDNF Family Neurotrophic Factor Signaling: Four Masters, One Servant?Mol. Cell. Neurosci.13, 313–325 (1999).doi: 10.1006/mcne.1999.0754
  4. Baloh, R. H., Enomoto, H., Johnson, E. M. & Milbrandt, J. The GDNF family ligands and receptors — implications for neural development.Curr. Opin. Neurobiol.10, 103–110 (2000).doi: 10.1016/s0959-4388(99)00048-3
  5. Akerud, P., Alberch, J., Eketjäll, S., Wagner, J. & Arenas, E. Differential effects of glial cell line-derived neurotrophic factor and neurturin on developing and adult substantia nigra dopaminergic neurons.J. Neurochem.73, 70–78 (1999).doi: 10.1046/j.1471-4159.1999.0730070.x
  6. Parsadanian, A., Pan, Y., Li, W., Myckatyn, T. M. & Brakefield, D. Astrocyte-derived transgene GDNF promotes complete and long-term survival of adult facial motoneurons following avulsion and differentially regulates the expression of transcription factors of AP-1 and ATF/CREB families.Exp. Neurol.200, 26–37 (2006).doi: 10.1016/j.expneurol.2006.01.014
  7. Boku, S.et al.GDNF facilitates differentiation of the adult dentate gyrus-derived neural precursor cells into astrocytes via STAT3.Biochem. Biophys. Res. Commun.434, 779–784 (2013).doi.org/10.1016/j.bbrc.2013.04.011
  8. Costantini, F. & Shakya, R. GDNF/Ret signaling and the development of the kidney.BioEssays News Rev. Mol. Cell. Dev. Biol.28, 117–127 (2006).doi.org/10.1002/bies.20357
  9. Hofmann, M.-C. Gdnf signaling pathways within the mammalian spermatogonial stem cell niche.Mol. Cell. Endocrinol.288, 95–103 (2008).doi: 10.1016/j.mce.2008.04.012
  10. Granholm, A.-C.et al.Glial Cell Line-Derived Neurotrophic Factor Is Essential for Postnatal Survival of Midbrain Dopamine Neurons.J. Neurosci.20, 3182–3190 (2000).doi.org/10.1523/JNEUROSCI.20-09-03182.2000
  11. Perrier, A. L.et al.Derivation of midbrain dopamine neurons from human embryonic stem cells.Proc. Natl. Acad. Sci. U. S. A.101, 12543–12548 (2004).doi.org/10.1073/pnas.0404700101
  12. Brennand, K.et al.Modeling schizophrenia using hiPSC neurons.Nature473, 221–225 (2011).doi: 10.1038/nature09915
  13. Kriks, S.et al.Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease.Nature480, 547–551 (2011).doi.org/10.1038/nature10648
  14. 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
  15. Li, X.-J.et al.Specification of motoneurons from human embryonic stem cells.Nat. Biotechnol.23, 215–221 (2005).doi: 10.1038/nbt1063
  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. McComish, S. F. & Caldwell, M. A. Generation of defined neural populations from pluripotent stem cells.Philos. Trans. R. Soc. B Biol. Sci.373, 20170214 (2018).doi: 10.1098/rstb.2017.0214
  18. Jacob, F.et al.Human Pluripotent Stem Cell-Derived Neural Cells and Brain Organoids Reveal SARS-CoV-2 Neurotropism Predominates in Choroid Plexus Epithelium.Cell Stem Cell27, 937-950.e9 (2020).doi: 10.1016/j.stem.2020.09.016
  19. Krencik, R., Weick, J. P., Liu, Y., Zhang, Z. & Zhang, S.-C. Specification of Transplantable Astroglial Subtypes from Human Pluripotent Stem Cells.Nat. Biotechnol.29, 528–534 (2011).doi: 10.1038/nbt.1877
  20. Serio, A.et al.Astrocyte pathology and the absence of non-cell autonomy in an induced pluripotent stem cell model of TDP-43 proteinopathy.Proc. Natl. Acad. Sci. U. S. A.110, 4697–4702 (2013).doi.org/10.1073/pnas.1300398110
  21. Gunhanlar, N.et al.A simplified protocol for differentiation of electrophysiologically mature neuronal networks from human induced pluripotent stem cells.Mol. Psychiatry23, 1336–1344 (2018).doi.org/10.1038/mp.2017.56
  22. Rocha, S. M., Cristovão, A. C., Campos, F. L., Fonseca, C. P. & Baltazar, G. Astrocyte-derived GDNF is a potent inhibitor of microglial activation.Neurobiol. Dis.47, 407–415 (2012).doi: 10.1016/j.nbd.2012.04.014
  23. Ibáñez, C. F. Emerging themes in structural biology of neurotrophic factors.Trends Neurosci.21, 438–444 (1998).doi: 10.1016/s0166-2236(98)01266-1
  24. Takahashi, M. The GDNF/RET signaling pathway and human diseases.Cytokine Growth Factor Rev.12, 361–373 (2001).doi: 10.1016/s1359-6101(01)00012-0
  25. Hsieh, J. C. & Penn, R. D. Chapter 44 – Infusion Therapy for Movement Disorders. inNeuromodulation(eds. Krames, E. S., Peckham, P. H. & Rezai, A. R.) 561–570 (Academic Press, 2009).doi:10.1016/B978-0-12-374248-3.00045-8.
  26. Manfredsson, F. P.et al.The Future of GDNF in Parkinson’s Disease.Front. Aging Neurosci.12,(2020). doi.org/10.3389/fnagi.2020.593572
  27. Barker, R. A.et al.GDNF and Parkinson’s Disease: Where Next? A Summary from a Recent Workshop.J. Park. Dis.10, 875–891.doi: 10.3233/JPD-202004