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Human GDNF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human GDNF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.


Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)



Data Example

Human GDNF ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: GDNF

GDNF (Glial Cell Line-derived Neurotrophic Factor) is a secreted protein that promotes the survival of motoneurons, midbrain dopaminergic neurons, Purkinje cells, and sympathetic neurons. It is produced by Sertoli cells, type 1 astrocytes, Schwann cells, neurons, pinealocytes, and skeletal muscle cells. GDNF binding to GFR alpha 1 induces the recruitment of Ret, NCAM-1/CD56, various integrins, Syndecan-3, or N-Cadherin. GDNF-based therapies show promise in several neurodegenerative disorders, particularly Parkinson’s disease.

Long Name:
Glial Cell line-derived Growth Factor
Entrez Gene IDs:
2668 (Human); 25453 (Rat)
Alternate Names:
Astrocyte-derived trophic factor; ATF; ATF1; ATF2; GDNF; glial cell derived neurotrophic factor; glial cell line derived neurotrophic factor; glial cell line-derived neurotrophic factor; HFB1-GDNF; hGDNF; HSCR3

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.


Citations for Human GDNF DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

27 Citations: Showing 1 - 10
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  1. Serum nerve growth factor beta, brain- and glial-derived neurotrophic factor levels and psychopathology in unmedicated patients with schizophrenia
    Authors: CS Chu, CL Chu, CC Wu, T Lu
    J Chin Med Assoc, 2018;0(0):.
    Species: Human
    Sample Types: Serum
  2. Melatonin antagonizes interleukin-18-mediated inhibition on neural stem cell proliferation and differentiation
    Authors: Z Li, X Li, MTV Chan, WKK Wu, D Tan, J Shen
    J. Cell. Mol. Med., 2017;0(0):.
    Species: Rat
    Sample Types: Cell Culture Supernates
  3. Elevated glutamate and lactate predict brain death after severe head trauma
    Authors: MA Stefani, R Modkovski, G Hansel, ER Zimmer, A Kopczynski, AP Muller, NR Strogulski, MS Rodolphi, RK Carteri, AP Schmidt, JP Oses, DH Smith, LV Portela
    Ann Clin Transl Neurol, 2017;4(6):392-402.
    Species: Human
    Sample Types: CSF
  4. A Novel Neuroprotective Small Molecule for Glial Cell Derived Neurotrophic Factor Induction and Photoreceptor Rescue
    Authors: P Baranov, H Lin, K McCabe, D Gale, S Cai, B Lieppman, D Morrow, P Lei, J Liao, M Young
    J Ocul Pharmacol Ther, 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. miRNA profiling of NurOwn�: mesenchymal stem cells secreting neurotrophic factors
    Authors: Y Gothelf, H Kaspi, N Abramov, R Aricha
    Stem Cell Res Ther, 2017;8(1):249.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Systemic injection of AAV9-GDNF provides modest functional improvements in the SOD1(G93A) ALS rat but has adverse side effects
    Authors: GM Thomsen, M Alkaslasi, JP Vit, G Lawless, M Godoy, G Gowing, O Shelest, CN Svendsen
    Gene Ther, 2017;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  7. Serum levels of neurotrophic factors in active toxoplasmic retinochoroiditis
    Authors: Cynthia Azeredo Cordeiro
    Braz J Infect Dis, 2016;0(0):.
    Species: Human
    Sample Types: Serum
  8. NG2 expression in microglial cells affects the expression of neurotrophic and proinflammatory factors by regulating FAK phosphorylation
    Authors: Lie Zhu
    Sci Rep, 2016;6(0):27983.
    Species: Mouse
    Sample Types: Serum
  9. Age and lesion-induced increases of GDNF transgene expression in brain following intracerebral injections of DNA nanoparticles.
    Authors: Yurek D, Hasselrot U, Cass W, Sesenoglu-Laird O, Padegimas L, Cooper M
    Neuroscience, 2015;284(0):500-12.
    Species: Human
    Sample Types: Tissue Homogenates
  10. Transplantation of human adipose tissue-derived stem cells delays clinical onset and prolongs life span in ALS mouse model.
    Authors: Kim, Kwang S, Lee, Hong J, An, Jin, Kim, Yun B, Ra, Jung Cha, Lim, Inja, Kim, Seung U
    Cell Transplant, 2014;23(12):1585-97.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Repeated systemic administration of human adipose-derived stem cells attenuates overt diabetic nephropathy in rats.
    Authors: Zhang L, Li K, Liu X, Li D, Luo C, Fu B, Cui S, Zhu F, Zhao R, Chen X
    Stem Cells Dev, 2013;22(23):3074-86.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Non-viral gene therapy for GDNF production in RCS rat: the crucial role of the plasmid dose.
    Gene Ther., 2012;19(9):886-98.
    Species: Rat
    Sample Types: Cell Culture Supernates
  13. A Monoclonal Antibody-GDNF Fusion Protein Is Not Neuroprotective and Is Associated with Proliferative Pancreatic Lesions in Parkinsonian Monkeys.
    Authors: Ohshima-Hosoyama S, Simmons HA, Goecks N
    PLoS ONE, 2012;7(6):e39036.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Types: Tissue Homogenates
  14. Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.
    Authors: Jones JL, Anderson JM, Phuah CL, Fox EJ, Selmaj K, Margolin D, Lake SL, Palmer J, Thompson SJ, Wilkins A, Webber DJ, Compston DA, Coles AJ
    Brain, 2010;133(0):2232-47.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Lack of humoral immune response to the tetracycline (Tet) activator in rats injected intracranially with Tet-off rAAV vectors.
    Authors: Han Y, Chang QA, Virag T
    Gene Ther., 2010;17(5):616-25.
    Species: Rat
    Sample Types: Tissue Homogenates
  16. Glial cell line-derived neurotrophic factor-secreting genetically modified human bone marrow-derived mesenchymal stem cells promote recovery in a rat model of Parkinson's disease.
    Authors: Glavaski-Joksimovic A, Virag T, Mangatu TA
    J. Neurosci. Res., 2010;88(12):2669-81.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Pharmacokinetics and bioactivity of glial cell line-derived factor (GDNF) and neurturin (NTN) infused into the rat brain.
    Authors: Hadaczek P, Johnston L, Forsayeth J, Bankiewicz KS
    Neuropharmacology, 2010;58(7):1114-21.
    Species: Rat
    Sample Types: Tissue Homogenates
  18. Effect of nigrostriatal damage induced by 6-hydroxydopamine on the expression of glial cell line-derived neurotrophic factor in the striatum of the rat.
    Authors: Mertens B, Massie A, Michotte Y, Sarre S
    Neuroscience, 2009;162(1):148-54.
    Species: Rat
    Sample Types: Tissue Homogenates
  19. Response of aged parkinsonian monkeys to in vivo gene transfer of GDNF.
    Authors: Emborg ME, Moirano J, Raschke J, Bondarenko V, Zufferey R, Peng S, Ebert AD, Joers V, Roitberg B, Holden JE, Koprich J, Lipton J, Kordower JH, Aebischer P
    Neurobiol. Dis., 2009;36(2):303-11.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Types: Tissue Homogenates
  20. Sustained intraspinal delivery of neurotrophic factor encapsulated in biodegradable nanoparticles following contusive spinal cord injury.
    Authors: Wang YC, Wu YT, Huang HY, Lin HI, Lo LW, Tzeng SF, Yang CS
    Biomaterials, 2008;29(34):4546-53.
    Species: Human
    Sample Types: Recombinant Protein
  21. Direct muscle delivery of GDNF with human mesenchymal stem cells improves motor neuron survival and function in a rat model of familial ALS.
    Authors: Suzuki M, McHugh J, Tork C, Shelley B, Hayes A, Bellantuono I, Aebischer P, Svendsen CN
    Mol. Ther., 2008;16(12):2002-10.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Transient striatal delivery of GDNF via encapsulated cells leads to sustained behavioral improvement in a bilateral model of Parkinson disease.
    Authors: Sajadi A, Bensadoun JC, Schneider BL, Lo Bianco C, Aebischer P
    Neurobiol. Dis., 2006;22(1):119-29.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Neurotrophic factors in relapsing remitting and secondary progressive multiple sclerosis patients during interferon beta therapy.
    Authors: Caggiula M, Batocchi AP, Frisullo G, Angelucci F, Patanella AK, Sancricca C, Nociti V, Tonali PA, Mirabella M
    Clin. Immunol., 2005;118(1):77-82.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Prolonged biologically active transgene expression driven by HSV LAP2 in brain in vivo.
    Authors: Puskovic V, Wolfe D, Goss J, Huang S, Mata M, Glorioso JC, Fink DJ
    Mol. Ther., 2004;10(1):67-75.
    Species: Rat
    Sample Types: Tissue Homogenates
  25. Intracerebral transplantation of carotid body in rats with transient middle cerebral artery occlusion.
    Authors: Yu G, Xu L, Hadman M, Hess DC, Borlongan CV
    Brain Res., 2004;1015(1):50-6.
    Species: Rat
    Sample Types: Tissue Homogenates
  26. AF5, a CNS cell line immortalized with an N-terminal fragment of SV40 large T: growth, differentiation, genetic stability, and gene expression.
    Authors: Truckenmiller ME, Vawter MP, Zhang P, Conejero-Goldberg C, Dillon-Carter O, Morales N, Cheadle C, Becker KG, Freed WJ
    Exp. Neurol., 2002;175(0):318.
    Species: Rat
    Sample Types: Cell Culture Supernates
  27. Lentiviral vectors as a gene delivery system in the mouse midbrain: cellular and behavioral improvements in a 6-OHDA model of Parkinson's disease using GDNF.
    Authors: Bensadoun JC, Deglon N, Tseng JL, Ridet JL, Zurn AD, Aebischer P
    Exp. Neurol., 2000;164(1):15-24.
    Species: Human
    Sample Types: Tissue Homogenates


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Reviews for Human GDNF DuoSet ELISA

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By Anonymous on 07/19/2017
Sample Tested: Serum-free Cell Culture Media

We are using this kit to detect the release of GDNF in serum of animal after implant our cells

By Anonymous on 05/06/2016
Sample Tested: uman serum