Human TGF-beta 3 DuoSet ELISA Summary
* 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
Product Datasheets
Preparation and Storage
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.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
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Citations: Showing 1 - 10
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Topographically and Chemically Enhanced Textile Polycaprolactone Scaffolds for Tendon and Ligament Tissue Engineering
Authors: Bauer, B;Emonts, C;Pitts, J;Buhl, EM;Eschweiler, J;Hänsch, R;Betsch, M;Gries, T;Menzel, H;
Polymers
Species: Human
Sample Types: Cell Culture Supernates
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Physiologic Doses of TGF-? Improve the Composition of Engineered Articular Cartilage
Authors: Wang, T;Dogru, S;Dai, Z;Kim, SY;Vickers, NA;Albro, MB;
bioRxiv : the preprint server for biology
Species: Bovine
Sample Types: Tissue Homogenates
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Release of TGF-beta3 from Surface-Modified PCL Fiber Mats Triggers a Dose-Dependent Chondrogenic Differentiation of Human Mesenchymal Stromal Cells
Authors: L Berten-Sch, Y Roger, H Bunjes, A Hoffmann
Pharmaceutics, 2023-04-21;15(4):.
Species: Human
Sample Types: Complex Sample Type
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Mesenchymal stromal cells promote the drug resistance of gastrointestinal stromal tumors by activating the PI3K-AKT pathway via TGF-beta2
Authors: Y Zhao, Z Weng, X Zhou, Z Xu, B Cao, B Wang, J Li
Journal of Translational Medicine, 2023-03-25;21(1):219.
Species: Human
Sample Types: Cell Culture Supernates
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Effect of IL-10 Deficiency on TGFbeta Expression during Fatal Alphavirus Encephalomyelitis in C57Bl/6 Mice
Authors: NM Martin, DE Griffin
Viruses, 2022-08-16;14(8):.
Species: Mouse
Sample Types: Tissue Homogenates
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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, 2022-03-31;12(4):.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2021-09-01;53(0):101336.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2021-01-21;22(3):.
Species: Human
Sample Types: Cell Lysates
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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, 2020-12-13;587(0):240-251.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2020-08-14;6(1):179-190.
Species: Rat
Sample Types: Cell Culture Supernates
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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, 2019-11-07;229(0):115551.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2019-11-03;20(21):.
Species: Human
Sample Types: Tissue Homogenates
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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, 2019-04-13;120(0):1-8.
Species: Rat
Sample Types: Cell Culture Supernates
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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, 2018-02-15;0(0):1-11.
Species: Human
Sample Types: Collagen Scaffold
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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, 2017-07-27;2017(0):9542702.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2017-01-18;2017(0):2730472.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2016-12-08;0(0):.
Species: Mouse
Sample Types: Cell Lysates
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The differential expression of protease activated receptors contributes to functional differences between dark and fair keratinocytes
Authors: Meilang Xue
J. Dermatol. Sci, 2016-12-05;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2014-09-10;9(9):e106773.
Species: Human
Sample Types: Peritoneal Fluid
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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, 2014-02-18;111(9):E798-806.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2014-01-13;111(4):E465-73.
Species: Mouse
Sample Types: Tissue Homogenates
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TGF-beta signaling regulates neuronal C1q expression and developmental synaptic refinement.
Authors: Bialas A, Stevens B
Nat Neurosci, 2013-10-27;16(12):1773-82.
Species: Rat
Sample Types: Cell Culture Supernates
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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, 2013-07-15;22(1):186-95.
Species: Rabbit
Sample Types: Cell Culture Supernates
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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, 2012-12-03;33(1):97-107.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2008-08-18;27(54):6834-44.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2007-11-26;149(3):1015-25.
Species: Human
Sample Types: Tissue Homogenates
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Autologous stem cell regeneration in craniosynostosis.
Authors: Moioli EK, Clark PA, Sumner DR, Mao JJ
Bone, 2007-10-17;42(2):332-40.
Species: Rat
Sample Types: Cell Culture Supernates
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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., 2007-08-29;176(10):974-82.
Species: Mouse
Sample Types: BALF
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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., 2007-01-01;210(1):192-200.
Species: Human
Sample Types: Cell Culture Supernates
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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., 2006-08-04;281(40):29479-90.
Species: Rat
Sample Types: Cell Culture Supernates
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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., 2006-05-01;176(9):5637-43.
Species: Human
Sample Types: Tissue Homogenates
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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., 2006-03-30;35(2):252-9.
Species: Human
Sample Types: Cell Culture Supernates
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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., 2004-07-01;152(1):191-4.
Species: Human
Sample Types: Plasma
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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., 2004-05-15;64(10):3566-71.
Species: Rat
Sample Types: Cell Culture Supernates
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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., 2002-09-16;277(47):45547-57.
Species: Human
Sample Types: Cell Culture Supernates
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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., 2002-08-06;71(4):329-34.
Species: Human
Sample Types: Cell Culture Supernates
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Amazing kit! Very easy to use and clear instructions. The kit provided highly reproducible results over time.