Human Pro-BDNF DuoSet ELISA

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Human Pro BDNF ELISA Standard Curve
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Product Details
Citations (12)
Supplemental Products
Reviews (2)

Human Pro-BDNF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
78.1 - 5,000 pg/mL
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 Pro-BDNF. 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)

Scientific Data

Human Pro BDNF ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: BDNF

BDNF (Brain-Derived Neurotrophic Factor) is a secreted protein that regulates many aspects of neuronal development and function in the nervous system. It is produced as proBDNF in response to neuronal activity or inflammatory stimulation and is then cleaved before associating into a homodimer. It is expressed throughout the nervous system as well as by fibroblasts, megakaryocytes/platelets, and smooth muscle cells. BDNF regulates neural stem cell survival and differentiation, axon/dendrite differentiation, synapse formation and maturation, and refinement of developing circuits. It is also a key regulator of synaptic plasticity and late-phase long-term potentiation. BDNF signals through the TrkB and NGF R/TNFRSF16 receptors. Secreted proBDNF can bind to NGF R/TNFRSF16 and induce long-term depression and cell apoptosis.

Long Name:
Brain-derived Neurotrophic Factor
Entrez Gene IDs:
627 (Human); 12064 (Mouse); 24225 (Rat)
Alternate Names:
Abrineurin; ANON2; BDNF; brain-derived neurotrophic factor; BULN2; MGC34632; Neurotrophin

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 Pro-BDNF 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.

12 Citations: Showing 1 - 10
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  1. Elevated Plasma Levels of Mature Brain-Derived Neurotrophic Factor in Major Depressive Disorder Patients with Higher Suicidal Ideation
    Authors: Li, H;Zhao, M;Jiang, C;Zhao, H;Wu, C;Li, Y;Zhang, S;Xu, P;Mou, T;Xu, Y;Huang, M;
    Brain sciences
    Species: Human
    Sample Types: Plasma
  2. The relationship between the brain-derived neurotrophic factor and neurocognitive response to physical exercise in individuals with schizophrenia
    Authors: Bang-Kittilsen, G;Egeland, J;Ueland, T;Andersen, E;Bigseth, TT;Holmen, TL;Mordal, J;Holst, R;Engh, JA;
    Species: Human
    Sample Types: Platelet-Poor Plasma
  3. The role of acute changes in mBDNF, cortisol and pro-BDNF in predicting cognitive performance in old age
    Authors: Nilsson, J;Ekblom, M;Moberg, M;Lövdén, M;
    Scientific reports
    Species: Human
    Sample Types: Plasma
  4. alpha7 Nicotinic acetylcholine receptor potentiation downregulates chemotherapy-induced inflammatory overactivation by overlapping intracellular mechanisms
    Authors: I Marmouzi, S Myers, DJ Buck, RL Davis, HR Arias
    The international journal of biochemistry & cell biology, 2023-03-24;158(0):106405.
    Species: Mouse
    Sample Types: Tissue Lysates
  5. BDNF, proBDNF and IGF-1 serum levels in na�ve and medicated subjects with autism
    Authors: MLA Robinson-A, B Michalski, B Vidal-Mart, LR Hernández, MW Santiesteb, M Fahnestock
    Scientific Reports, 2022-08-12;12(1):13768.
    Species: Human
    Sample Types: Serum
  6. BDNF and Pro-BDNF in Amyotrophic Lateral Sclerosis: A New Perspective for Biomarkers of Neurodegeneration
    Authors: G Riolo, C Ricci, N De Angelis, C Marzocchi, G Guerrera, G Borsellino, F Giannini, S Battistini
    Brain sciences, 2022-05-09;12(5):.
    Species: Human
    Sample Types: Serum
  7. ProBDNF as an Indicator of Improvement among Women with Depressive Episodes
    Authors: W Zwoli?ska, M Skibinska, A S?opie?, M Dmitrzak-W
    Metabolites, 2022-04-16;12(4):.
    Species: Human
    Sample Types: Serum
  8. Increased ratio of mature BDNF to precursor-BDNF in patients with major depressive disorder with severe anhedonia
    Authors: C Wu, J Lu, S Lu, M Huang, Y Xu
    J Psychiatr Res, 2020-05-12;126(0):92-97.
    Species: Human
    Sample Types: Plasma
  9. N-Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures
    Authors: J Benicky, M Sanda, Z Brnakova K, R Goldman
    J. Biol. Chem., 2019-09-26;0(0):.
    Species: Rat
    Sample Types: Cell Culture Supernates
  10. Learning and memory deficits and alzheimer's disease-like changes in mice after chronic exposure to microcystin-LR
    Authors: J Wang, Y Chen, C Zhang, Z Xiang, J Ding, X Han
    J. Hazard. Mater., 2019-03-26;373(0):504-518.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. The serum protein levels of the tPA-BDNF pathway are implicated in depression and antidepressant treatment
    Authors: H Jiang, S Chen, C Li, N Lu, Y Yue, Y Yin, Y Zhang, X Zhi, D Zhang, Y Yuan
    Transl Psychiatry, 2017-04-04;7(4):e1079.
    Species: Human
    Sample Types: Serum
  12. Serum levels of neurotrophic factors in active toxoplasmic retinochoroiditis
    Authors: Cynthia Azeredo Cordeiro
    Braz J Infect Dis, 2016-12-05;0(0):.
    Species: Human
    Sample Types: Serum


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Reviews for Human Pro-BDNF DuoSet ELISA

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Human Pro-BDNF DuoSet ELISA
By Anonymous on 11/15/2023
Sample Tested: Plasma

Human Pro-BDNF DuoSet ELISA
By Anonymous on 10/24/2019
Sample Tested: EDTA Plasma

Used in EDTA Plasma diluted 1:10 in Reagent Diluent. Replicates are very good, samples well into detection range.