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Human TGF-beta 3 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 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 TGF-beta3. 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 TGF-beta 3 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: TGF-beta 3

Transforming Growth Factor Beta 1, 2, and 3 (TGF-beta 1, TGF-beta 2, and TGF-beta 3) are highly pleiotropic cytokines that virtually all cell types secrete. TGF-beta molecules are proposed to act as cellular switches that regulate processes such as immune function, proliferation, and epithelial-mesenchymal transition. Targeted deletions of these genes in mice show that each TGF-beta isoform has some non-redundant functions: TGF-beta 1 is involved in hematopoiesis and endothelial differentiation; TGF-beta 2 affects development of cardiac, lung, craniofacial, limb, eye, ear, and urogenital systems; and TGF-beta 3 influences palatogenesis and pulmonary development. The full range of in vitro biological activities of TGF-beta 5 has not yet been explored. However, TGF-beta 1, TGF-beta 2, TGF-beta 3, and TGF-beta 5 have been found to be largely interchangeable in an inhibitory bioassay, and it is anticipated that TGF-beta 5 will show a spectrum of activities similar to the other TGF-beta family members. To date, the production of TGF-beta 5 has only been demonstrated in Xenopus.

TGF-beta ligands are initially synthesized as precursor proteins that undergo proteolytic cleavage. The mature segments form active ligand dimers via a disulfide-rich core consisting of the characteristic 'cysteine knot'. TGF-beta signaling begins with binding to a complex of the accessory receptor betaglycan (also known as TGF-beta RIII) and a type II serine/threonine kinase receptor termed TGF-beta RII. This receptor then phosphorylates and activates a type I serine/threonine kinase receptor, either ALK-1 or TGF-beta RI (also called ALK-5). The activated type I receptor phosphorylates and activates Smad proteins that regulate transcription. Use of other signaling pathways that are Smad-independent allows for distinct actions observed in response to TGF-beta in different contexts.

Long Name:
Transforming Growth Factor beta 3
Entrez Gene IDs:
7043 (Human); 21809 (Mouse); 25717 (Rat)
Alternate Names:
ARVD; ARVD1; FLJ16571; LDS5; RNHF; TGFB3; TGFbeta 3; TGF-beta 3; TGF-beta3; TGF-beta-3; transforming growth factor beta-3; transforming growth factor, beta 3

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 TGF-beta 3 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.

32 Citations: Showing 1 - 10
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  1. Effect of IL-10 Deficiency on TGFbeta Expression during Fatal Alphavirus Encephalomyelitis in C57Bl/6 Mice
    Authors: NM Martin, DE Griffin
    Viruses, 2022;14(8):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. Cytokine Profile and Anti-Inflammatory Activity of a Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells (PRS CK STORM)
    Authors: JP Lapuente, A Blázquez-M, J Marco-Brua, G Gómez, P Desportes, J Sanz, P Fernández, M García-Gil, F Bermejo, JV San Martín, A Algaba, JC De Gregori, D Lapuente, A De Gregori, B Lapuente, MV Andrés, A Anel
    Biomolecules, 2022;12(4):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Regulation of Fibroblast Activation Protein by Transforming Growth Factor Beta-1 in Glioblastoma Microenvironment
    Authors: E Krepela, Z Vanickova, P Hrabal, M Zubal, B Chmielova, E Balaziova, P Vymola, I Matrasova, P Busek, A Sedo
    International Journal of Molecular Sciences, 2021;22(3):.
    Species: Human
    Sample Types: Cell Lysates
  4. A 3D-Bioprinted dual growth factor-releasing intervertebral disc scaffold induces nucleus pulposus and annulus fibrosus reconstruction
    Authors: B Sun, M Lian, Y Han, X Mo, W Jiang, Z Qiao, K Dai
    Bioact Mater, 2021;6(1):179-190.
    Species: Rat
    Sample Types: Cell Culture Supernates
  5. Latent TGFbeta-binding proteins regulate UCP1 expression and function via TGFbeta2
    Authors: D Halbgebaue, J Roos, JB Funcke, H Neubauer, BS Hamilton, E Simon, EZ Amri, KM Debatin, M Wabitsch, P Fischer-Po, D Tews
    Molecular Metabolism, 2021;53(0):101336.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Formation of alginate microspheres prepared by optimized microfluidics parameters for high encapsulation of bioactive molecules
    Authors: LM Caballero, S Duchi, C Onofrillo, CD O'Connell, C Di Bella, SE Moulton
    Journal of colloid and interface science, 2020;587(0):240-251.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Promoted chondrogenesis of hMCSs with controlled release of TGF-beta3 via microfluidics synthesized alginate nanogels
    Authors: Z Mahmoudi, J Mohammadne, S Razavi Baz, A Abouei Meh, M Saidijam, R Dinarvand, M Ebrahimi W, M Soleimani
    Carbohydr Polym, 2020;229(0):115551.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Transforming Growth Factor Beta 3-Loaded Decellularized Equine Tendon Matrix for Orthopedic Tissue Engineering
    Authors: SP Roth, W Brehm, C Gro beta, P Scheibe, S Schubert, J Burk
    Int J Mol Sci, 2019;20(21):.
    Species: Human
    Sample Types: Tissue Homogenates
  9. Synergistic effects of recombinant Lentiviral-mediated BMP2 and TGF-beta3 on the osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro
    Authors: W He, L Chen, Y Huang, Z Xu, W Xu, N Ding, J Chen
    Cytokine, 2019;120(0):1-8.
    Species: Rat
    Sample Types: Cell Culture Supernates
  10. Restoring tracheal defects in a rabbit model with tissue engineered patches based on TGF-?3-encapsulating electrospun poly(l-lactic acid-co-?-caprolactone)/collagen scaffolds
    Authors: H Jing, B Gao, M Gao, H Yin, X Mo, X Zhang, K Luo, B Feng, W Fu, J Wang, W Zhang, M Yin, Z Zhu, X He, J Zheng
    Artif Cells Nanomed Biotechnol, 2018;0(0):1-11.
    Species: Human
    Sample Types: Collagen Scaffold
  11. Signal Factors Secreted by 2D and Spheroid Mesenchymal Stem Cells and by Cocultures of Mesenchymal Stem Cells Derived Microvesicles and Retinal Photoreceptor Neurons
    Authors: L Xie, M Mao, L Zhou, L Zhang, B Jiang
    Stem Cells Int, 2017;2017(0):2730472.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Paracrine Potential of the Human Adipose Tissue-Derived Stem Cells to Modulate Balance between Matrix Metalloproteinases and Their Inhibitors in the Osteoarthritic Cartilage In Vitro
    Authors: J Denkovskij, E Bagdonas, I Kusleviciu, Z Mackiewicz, A Unguryte, N Porvanecka, S Fleury, A Venalis, C Jorgensen, E Bernotiene
    Stem Cells Int, 2017;2017(0):9542702.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. The differential expression of protease activated receptors contributes to functional differences between dark and fair keratinocytes
    Authors: Meilang Xue
    J. Dermatol. Sci, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Neuronal Protein 3.1 Deficiency Leads to Reduced Cutaneous Scar Collagen Deposition and Tensile Strength due to Impaired Transforming Growth Factor-?1 to -?3 Translation
    Authors: Tao Cheng
    Am. J. Pathol, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
  15. Regenerating cartilages by engineered ASCs: prolonged TGF-beta3/BMP-6 expression improved articular cartilage formation and restored zonal structure.
    Authors: Lu C, Yeh T, Yeh C, Fang Y, Sung L, Lin S, Yen T, Chang Y, Hu Y
    Mol Ther, 2014;22(1):186-95.
    Species: Rabbit
    Sample Types: Cell Culture Supernates
  16. Thymocyte apoptosis drives the intrathymic generation of regulatory T cells.
    Authors: Konkel J, Jin W, Abbatiello B, Grainger J, Chen W
    Proc Natl Acad Sci U S A, 2014;111(4):E465-73.
    Species: Mouse
    Sample Types: Tissue Homogenates
  17. Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage.
    Authors: Brunger J, Huynh N, Guenther C, Perez-Pinera P, Moutos F, Sanchez-Adams J, Gersbach C, Guilak F
    Proc Natl Acad Sci U S A, 2014;111(9):E798-806.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Interaction with colon cancer cells hyperactivates TGF-beta signaling in cancer-associated fibroblasts.
    Authors: Hawinkels L, Paauwe M, Verspaget H, Wiercinska E, van der Zon J, van der Ploeg K, Koelink P, Lindeman J, Mesker W, ten Dijke P, Sier C
    Oncogene, 2014;33(1):97-107.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. The peritoneum is both a source and target of TGF-beta in women with endometriosis.
    Authors: Young V, Brown J, Saunders P, Duncan W, Horne A
    PLoS ONE, 2014;9(9):e106773.
    Species: Human
    Sample Types: Peritoneal Fluid
  20. TGF-beta signaling regulates neuronal C1q expression and developmental synaptic refinement.
    Authors: Bialas A, Stevens B
    Nat Neurosci, 2013;16(12):1773-82.
    Species: Rat
    Sample Types: Cell Culture Supernates
  21. Glioblastoma-secreted factors induce IGFBP7 and angiogenesis by modulating Smad-2-dependent TGF-beta signaling.
    Authors: Pen A, Moreno MJ, Durocher Y, Deb-Rinker P, Stanimirovic DB
    Oncogene, 2008;27(54):6834-44.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Autologous stem cell regeneration in craniosynostosis.
    Authors: Moioli EK, Clark PA, Sumner DR, Mao JJ
    Bone, 2007;42(2):332-40.
    Species: Rat
    Sample Types: Cell Culture Supernates
  23. Transforming growth factor-beta1 suppresses airway hyperresponsiveness in allergic airway disease.
    Authors: Alcorn JF, Rinaldi LM, Jaffe EF, van Loon M, Bates JH, Janssen-Heininger YM, Irvin CG
    Am. J. Respir. Crit. Care Med., 2007;176(10):974-82.
    Species: Mouse
    Sample Types: BALF
  24. Opposite regulation of transforming growth factors-beta2 and -beta3 expression in the human endometrium.
    Authors: Gaide Chevronnay HP, Cornet PB, Delvaux D, Lemoine P, Courtoy PJ, Henriet P, Marbaix E
    Endocrinology, 2007;149(3):1015-25.
    Species: Human
    Sample Types: Tissue Homogenates
  25. Interferon-gamma differentially regulates TGF-beta1 and TGF-beta2 expression in human retinal pigment epithelial cells through JAK-STAT pathway.
    Authors: Nagineni CN, Cherukuri KS, Kutty V, Detrick B, Hooks JJ
    J. Cell. Physiol., 2007;210(1):192-200.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. Activation of transforming growth factor-beta by the integrin alphavbeta8 delays epithelial wound closure.
    Authors: Neurohr C, Nishimura SL, Sheppard D
    Am. J. Respir. Cell Mol. Biol., 2006;35(2):252-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Requirement of Smad3 and CREB-1 in mediating transforming growth factor-beta (TGF beta) induction of TGF beta 3 secretion.
    Authors: Liu G, Ding W, Neiman J, Mulder KM
    J. Biol. Chem., 2006;281(40):29479-90.
    Species: Rat
    Sample Types: Cell Culture Supernates
  28. Tumor-derived TGFbeta-1 induces dendritic cell apoptosis in the sentinel lymph node.
    Authors: Ito M, Minamiya Y, Kawai H, Saito S, Saito H, Nakagawa T, Imai K, Hirokawa M, Ogawa J
    J. Immunol., 2006;176(9):5637-43.
    Species: Human
    Sample Types: Tissue Homogenates
  29. Increased blood plasma concentrations of TGF-beta isoforms after treatment with intravenous immunoglobulins (i.v.IG) in patients with multiple sclerosis.
    Authors: Reinhold D, Perlov E, Schrecke K, Kekow J, Brune T, Sailer M
    J. Neuroimmunol., 2004;152(1):191-4.
    Species: Human
    Sample Types: Plasma
  30. The selective estrogen receptor modulator arzoxifene and the rexinoid LG100268 cooperate to promote transforming growth factor beta-dependent apoptosis in breast cancer.
    Authors: Rendi MH, Suh N, Lamph WW, Krajewski S, Reed JC, Heyman RA, Berchuck A, Liby K, Risingsong R, Royce DB, Williams CR, Sporn MB
    Cancer Res., 2004;64(10):3566-71.
    Species: Rat
    Sample Types: Cell Culture Supernates
  31. Progesterone upregulates TGF-b isoforms (b1, b2, and b3) expression in normal human osteoblast-like cells.
    Authors: Luo XH, Liao EY, Su X
    Calcif. Tissue Int., 2002;71(4):329-34.
    Species: Human
    Sample Types: Cell Culture Supernates
  32. Transforming growth factor-beta -Smad signaling pathway cooperates with NF-kappa B to mediate nontypeable Haemophilus influenzae-induced MUC2 mucin transcription.
    Authors: Jono H, Shuto T, Xu H, Kai H, Lim DJ, Gum JR, Kim YS, Yamaoka S, Feng XH, Li JD
    J. Biol. Chem., 2002;277(47):45547-57.
    Species: Human
    Sample Types: Cell Culture Supernates


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Human TGF-beta 3 DuoSet ELISA
By Anonymous on 04/09/2021
Sample Tested: TGF-beta 3 released from synthetic hydrogels

Amazing kit! Very easy to use and clear instructions. The kit provided highly reproducible results over time.

Human TGF-beta 3 DuoSet ELISA
By Anonymous on 04/02/2020
Sample Tested: Serum and Plasma

We used this kit for the quantification of TGF b3 in human serum and plasma. Works very well and well-described protocol.