Human TGF-alpha DuoSet ELISA

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Human TGF-alpha ELISA Standard Curve
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Product Details
Citations (24)
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Human TGF-alpha DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
7.8 - 500 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-alpha. 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-alpha 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-alpha

TGF-alpha is a member of the EGF family of cytokines that are synthesized as transmembrane precursors with soluble forms released by proteolytic cleavage. Membrane-bound proTGF-alpha is biologically active and plays a role in mediation of cell-cell adhesion and in juxtacrine stimulation of adjacent cells. Expression of TGF-alpha is widespread in tumors and transformed cells. TGF-alpha is also expressed in normal tissues during embryogenesis and in adult tissues, including pituitary, brain, keratinocytes and macrophages. TGF-alpha binds and activates the EGF receptor. It induces fibroblast proliferation, epithelial development, keratinocyte migration, and angiogenesis.

Long Name:
Transforming Growth Factor alpha
Entrez Gene IDs:
7039 (Human)
Alternate Names:
protransforming growth factor alpha; TFGA; TGFA; TGFalpha; TGF-alpha; transforming growth factor, alpha; transforming growth factor-alpha

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-alpha 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.

24 Citations: Showing 1 - 10
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  1. Role of iRhoms 1 and 2 in Endochondral Ossification
    Authors: R Fang, C Haxaire, M Otero, S Lessard, G Weskamp, DR McIlwain, TW Mak, SF Lichtentha, CP Blobel
    Int J Mol Sci, 2020-11-19;21(22):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Efficient blockade of locally reciprocated tumor-macrophage signaling using a TAM-avid nanotherapy
    Authors: SJ Wang, R Li, TSC Ng, G Luthria, MJ Oudin, M Prytyskach, RH Kohler, R Weissleder, DA Lauffenbur, MA Miller
    Sci Adv, 2020-05-22;6(21):eaaz8521.
    Species: Human
    Sample Types: Cell Lysates
  3. HER-family ligands promote acquired resistance to trastuzumab in gastric cancer
    Authors: A Sampera, FJ Sánchez-Ma, O Arpí, L Visa, M Iglesias, S Menéndez, É Gaye, A Dalmases, S Clavé, M Gelabert-B, TT Poulsen, M Kragh, B Bellosillo, J Albanell, A Rovira, C Montagut
    Mol. Cancer Ther., 2019-09-04;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Human rhomboid family-1 modulates clathrin coated vesicle-dependent pro-transforming growth factor ? membrane trafficking to promote breast cancer progression
    Authors: J Li, TR Bai, S Gao, Z Zhou, XM Peng, LS Zhang, DL Dou, ZS Zhang, LY Li
    EBioMedicine, 2018-09-29;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. p63 is a key regulator of iRHOM2 signalling in the keratinocyte stress response
    Authors: P Arcidiacon, CM Webb, MA Brooke, H Zhou, PJ Delaney, KE Ng, DC Blaydon, A Tinker, DP Kelsell, A Chikh
    Nat Commun, 2018-03-09;9(1):1021.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Functional Genomics Approach Identifies Novel Signaling Regulators of TGF? Ectodomain Shedding
    Authors: JL Wilson, E Kefalogian, L Stopfer, C Harrison, VS Sabbisetti, E Fraenkel, DA Lauffenbur, A Herrlich
    Mol. Cancer Res., 2017-10-10;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Five-weekly S-1 plus cisplatin therapy combined with trastuzumab therapy in HER2-positive gastric cancer: a phase II trial and biomarker study (WJOG7212G)
    Authors: Y Miura, Y Sukawa, S Hironaka, M Mori, K Nishikawa, S Tokunaga, H Okuda, T Sakamoto, K Taku, K Nishikawa, T Moriwaki, Y Negoro, Y Kimura, K Uchino, K Shinozaki, H Shinozaki, N Musha, H Yoshiyama, T Tsuda, Y Miyata, N Sugimoto, T Shirakawa, M Ito, K Yonesaka, K Yoshimura, N Boku, K Nosho, T Takano, I Hyodo
    Gastric Cancer, 2017-05-11;0(0):.
    Species: Human
    Sample Types: Serum
  8. Prognostic value of rare and complex mutations in EGFR and serum levels of soluble EGFR and its ligands in Iranian non-small cell lung carcinoma patients
    Authors: Seyyed Mortaza Haghgoo
    Clin. Biochem, 2016-12-05;0(0):.
    Species: Human
    Sample Types: Serum
  9. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016-08-18;1(13):.
    Species: Human
    Sample Types: Urine
  10. RAF inhibitors that evade paradoxical MAPK pathway activation.
    Authors: Zhang C, Spevak W, Zhang Y, Burton E, Ma Y, Habets G, Zhang J, Lin J, Ewing T, Matusow B, Tsang G, Marimuthu A, Cho H, Wu G, Wang W, Fong D, Nguyen H, Shi S, Womack P, Nespi M, Shellooe R, Carias H, Powell B, Light E, Sanftner L, Walters J, Tsai J, West B, Visor G, Rezaei H, Lin P, Nolop K, Ibrahim P, Hirth P, Bollag G
    Nature, 2015-10-14;526(7574):583-6.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. Smooth muscle cells relay acute pulmonary inflammation via distinct ADAM17/ErbB axes.
    Authors: Dreymueller D, Martin C, Schumacher J, Groth E, Boehm J, Reiss L, Uhlig S, Ludwig A
    J Immunol, 2013-12-16;192(2):722-31.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. A disintegrin and metalloproteinase 17 (ADAM17) and epidermal growth factor receptor (EGFR) signaling drive the epithelial response to Staphylococcus aureus toxic shock syndrome toxin-1 (TSST-1).
    Authors: Breshears L, Schlievert P, Peterson M
    J Biol Chem, 2012-07-25;287(39):32578-87.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Anti-Tumour Effects of a Specific Anti-ADAM17 Antibody in an Ovarian Cancer Model In Vivo.
    Authors: Richards FM, Tape CJ, Jodrell DI, Murphy G
    PLoS ONE, 2012-07-11;7(7):e40597.
    Species: Human
    Sample Types: Tissue Homogenates
  14. LPA-producing enzyme PA-PLA(1)alpha regulates hair follicle development by modulating EGFR signalling.
    Authors: Inoue A, Arima N, Ishiguro J, Prestwich GD, Arai H, Aoki J
    EMBO J., 2011-08-19;30(20):4248-60.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  15. The EGFR ligands amphiregulin and heparin-binding egf-like growth factor promote peritoneal carcinomatosis in CXCR4-expressing gastric cancer.
    Authors: Yasumoto K, Yamada T, Kawashima A, Wang W, Li Q, Donev IS, Tacheuchi S, Mouri H, Yamashita K, Ohtsubo K, Yano S
    Clin. Cancer Res., 2011-04-11;17(11):3619-30.
    Species: Human
    Sample Types: Complex Sample Type
  16. Ectodomain shedding of EGFR ligands and TNFR1 dictates hepatocyte apoptosis during fulminant hepatitis in mice.
    Authors: Murthy A, Defamie V, Smookler DS
    J. Clin. Invest., 2010-07-12;120(8):2731-44.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  17. Identification of biomarkers in human head and neck tumor cell lines that predict for in vitro sensitivity to gefitinib.
    Authors: Hickinson DM, Marshall GB, Beran GJ
    Clin Transl Sci, 2009-06-01;2(3):183-92.
    Species: Human
    Sample Types: Cell Lysates
  18. Amphiregulin is much more abundantly expressed than transforming growth factor-alpha and epidermal growth factor in human follicular fluid obtained from patients undergoing in vitro fertilization-embryo transfer.
    Authors: Inoue Y, Miyamoto S, Fukami T, Shirota K, Yotsumoto F, Kawarabayashi T
    Fertil. Steril., 2008-03-05;91(4):1035-41.
    Species: Human
    Sample Types: Follicular Fluid
  19. Validation of HB-EGF and amphiregulin as targets for human cancer therapy.
    Authors: Yotsumoto F, Yagi H, Suzuki SO, Oki E, Tsujioka H, Hachisuga T, Sonoda K, Kawarabayashi T, Mekada E, Miyamoto S
    Biochem. Biophys. Res. Commun., 2007-11-20;365(3):555-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Expression and role of EGFR ligands induced in airway cells by PM2.5 and its components.
    Authors: Rumelhard M, Ramgolam K, Hamel R, Marano F, Baeza-Squiban A
    Eur. Respir. J., 2007-09-05;30(6):1064-73.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. In-frame deletion in the EGF receptor alters kinase inhibition by gefitinib.
    Authors: Sakai K, Yokote H, Murakami-Murofushi K, Tamura T, Saijo N, Nishio K
    Biochem. J., 2006-08-01;397(3):537-43.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Targeting ADAM-mediated ligand cleavage to inhibit HER3 and EGFR pathways in non-small cell lung cancer.
    Authors: Zhou BB, Peyton M, He B, Liu C, Girard L, Caudler E, Lo Y, Baribaud F, Mikami I, Reguart N, Yang G, Li Y, Yao W, Vaddi K, Gazdar AF, Friedman SM, Jablons DM, Newton RC, Fridman JS, Minna JD, Scherle PA
    Cancer Cell, 2006-07-01;10(1):39-50.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Cigarette smoke synergistically enhances respiratory mucin induction by proinflammatory stimuli.
    Authors: Baginski TK, Dabbagh K, Satjawatcharaphong C, Swinney DC
    Am. J. Respir. Cell Mol. Biol., 2006-03-16;35(2):165-74.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Heparin-binding EGF-like growth factor is a promising target for ovarian cancer therapy.
    Authors: Miyamoto S, Hirata M, Yamazaki A, Kageyama T, Hasuwa H, Mizushima H, Tanaka Y, Yagi H, Sonoda K, Kai M, Kanoh H, Nakano H, Mekada E
    Cancer Res., 2004-08-15;64(16):5720-7.
    Species: Human
    Sample Types: Ascites Fluid


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