Human Amphiregulin DuoSet ELISA

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Human Amphiregulin ELISA Standard Curve
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Citations (42)
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Human Amphiregulin DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 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 Amphiregulin (AR). 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 Amphiregulin 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: Amphiregulin

Amphiregulin (AR) is an EGF family growth factor that is released as a soluble protein following proteolytic cleavage of its transmembrane precursor. Amphiregulin is expressed by numerous carcinoma cell lines and epithelial cells of the colon, stomach, breast, ovary and kidney. It acts through ErbB family of receptor to stimulate the proliferation of keratinocytes, mammary epithelial cells, fibroblasts, astrocytes and glial cells. Amphiregulin can also inhibit the growth of certain tumor cells.

Entrez Gene IDs:
374 (Human); 11839 (Mouse)
Alternate Names:
Amphiregulin; AR; AREG; AREGB; Colorectum cell-derived growth factor; CRDGF; MGC13647; schwannoma-derived growth factor; SDGF

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 Amphiregulin 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.

42 Citations: Showing 1 - 10
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  1. Agricultural dust derived bacterial extracellular vesicle mediated inflammation is attenuated by DHA
    Authors: AJ Heires, D Samuelson, D Villageliu, TM Nordgren, DJ Romberger
    Scientific Reports, 2023-02-16;13(1):2767.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Airway basal cells show a dedifferentiated KRT17highPhenotype and promote fibrosis in idiopathic pulmonary fibrosis
    Authors: B Jaeger, JC Schupp, L Plappert, O Terwolbeck, N Artysh, G Kayser, P Engelhard, TS Adams, R Zweigerdt, H Kempf, S Lienenklau, W Garrels, I Nazarenko, D Jonigk, M Wygrecka, D Klatt, A Schambach, N Kaminski, A Prasse
    Nature Communications, 2022-09-26;13(1):5637.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Inflammatory and Proliferative Pathway Activation in Human Esophageal Myofibroblasts Treated with Acidic Bile Salts
    Authors: M Patankar, M Li, A Khalatbari, JD Castle, L Hu, C Zhang, A Shaker
    International Journal of Molecular Sciences, 2022-09-08;23(18):.
    Species: Human
    Sample Types: Cell Culture Supernatants
  4. Clinical and biological markers for predicting ARDS and outcome in septic patients
    Authors: J Villar, R Herrán-Mon, E González-H, M Prieto-Gon, A Ambrós, A Rodríguez-, A Muriel-Bom, R Solano, C Cuenca-Rub, A Vidal, C Flores, JM González-M, MI García-Lao, Genetics o
    Scientific Reports, 2021-11-22;11(1):22702.
    Species: Human
    Sample Types: Serum
  5. PD-L1 recruits phospholipase C and enhances tumorigenicity of lung tumors harboring mutant forms of EGFR
    Authors: S Ghosh, NB Nataraj, A Noronha, S Patkar, A Sekar, S Mukherjee, S Winograd-K, L Kramarski, A Verma, M Lindzen, DD Garcia, J Green, G Eisenberg, H Gil-Henn, A Basu, Y Lender, S Weiss, M Oren, M Lotem, B Geiger, E Ruppin, Y Yarden
    Cell Reports, 2021-05-25;35(8):109181.
    Species: Human
    Sample Types: Cell Cycle Supernates
  6. Targeting autocrine amphiregulin robustly and reproducibly inhibits ovarian cancer in a syngeneic model: roles for wildtype p53
    Authors: M Lindzen, S Ghosh, A Noronha, D Drago, NB Nataraj, O Leitner, S Carvalho, E Zmora, S Sapoznik, KB Shany, K Levanon, D Aderka, BS Ramírez, M Dahlhoff, I McNeish, Y Yarden
    Oncogene, 2021-04-30;0(0):.
    Species: Human
    Sample Types: Ascites Fluid
  7. Downregulation of epithelial DUOX1 in Chronic Obstructive Pulmonary Disease contributes to disease pathogenesis
    Authors: C Schiffers, C van de Wet, RA Bauer, A Habibovic, M Hristova, CM Dustin, S Lambrichts, PM Vacek, EF Wouters, NL Reynaert, A van der Vl
    JCI Insight, 2021-01-25;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. 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
  9. Mesenchymal Stem and Stromal Cells Harness Macrophage-Derived Amphiregulin to Maintain Tissue Homeostasis
    Authors: JH Ko, HJ Kim, HJ Jeong, HJ Lee, JY Oh
    Cell Rep, 2020-03-17;30(11):3806-3820.e6.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. 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
  11. Candidalysin activates innate epithelial immune responses via epidermal growth factor receptor
    Authors: J Ho, X Yang, SA Nikou, N Kichik, A Donkin, NO Ponde, JP Richardson, RL Gratacap, LS Archambaul, CP Zwirner, C Murciano, R Henley-Smi, S Thavaraj, CJ Tynan, SL Gaffen, B Hube, RT Wheeler, DL Moyes, JR Naglik
    Nat Commun, 2019-05-24;10(1):2297.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Innate Control of Tissue-Reparative Human Regulatory T Cells
    Authors: AJ Lam, KN MacDonald, AM Pesenacker, SC Juvet, KA Morishita, B Bressler, JG Pan, SS Sidhu, JD Rioux, MK Levings
    J. Immunol., 2019-03-08;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. AhR controls redox homeostasis and shapes the tumor microenvironment in BRCA1-associated breast cancer
    Authors: SP Kubli, C Bassi, C Roux, A Wakeham, C Göbl, W Zhou, SM Jafari, B Snow, L Jones, L Palomero, KL Thu, L Cassetta, D Soong, T Berger, P Ramachandr, SP Baniasadi, G Duncan, M Lindzen, Y Yarden, C Herranz, C Lazaro, MF Chu, J Haight, P Tinto, J Silvester, DW Cescon, A Petit, S Pettersson, JW Pollard, TW Mak, MA Pujana, P Cappello, C Gorrini
    Proc. Natl. Acad. Sci. U.S.A., 2019-02-07;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Effects of agricultural organic dusts on human lung-resident mesenchymal stem (stromal) cell function
    Authors: TM Nordgren, KL Bailey, AJ Heires, D Katafiasz, DJ Romberger
    Toxicol. Sci., 2018-04-01;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. ADAM17 is required for EGF-R-induced intestinal tumors via IL-6 trans-signaling
    Authors: S Schmidt, N Schumacher, J Schwarz, S Tangermann, L Kenner, M Schlederer, M Sibilia, M Linder, A Altendorf-, T Knösel, ES Gruber, G Oberhuber, J Bolik, A Rehman, A Sinha, J Lokau, P Arnold, AS Cabron, F Zunke, C Becker-Pau, A Preaudet, P Nguyen, J Huynh, S Afshar-Ste, AL Chand, J Westermann, PJ Dempsey, C Garbers, D Schmidt-Ar, P Rosenstiel, T Putoczki, M Ernst, S Rose-John
    J. Exp. Med., 2018-02-22;0(0):.
    Species: Mouse
    Sample Types: Tissue Culture Supernates
  16. 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
  17. Epidermal growth factor (EGF) receptor-ligand based molecular staging predicts prognosis in head and neck squamous cell carcinoma partly due to deregulated EGF- induced amphiregulin expression
    J Exp Clin Cancer Res, 2016-09-26;35(1):151.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016-08-18;1(13):.
    Species: Human
    Sample Types: Urine
  19. Local Epidermal Growth Factor Receptor Signaling Mediates the Systemic Pathogenic Effects of Staphylococcus aureus Toxic Shock Syndrome
    PLoS ONE, 2016-07-14;11(7):e0158969.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Targeting mTOR and AREG with everolimus, sunitinib and sorafenib in HPV-positive and -negative SCC.
    Authors: Aderhold C, Faber A, Umbreit C, Birk R, Weiss C, Sommer J, Hormann K, Schultz J
    Anticancer Res, 2015-04-01;35(4):1951-9.
    Species: Human
    Sample Types: Cell Lysates
  21. An integrative analysis of the tumorigenic role of TAZ in human non-small cell lung cancer.
    Authors: Noguchi S, Saito A, Horie M, Mikami Y, Suzuki H, Morishita Y, Ohshima M, Abiko Y, Mattsson J, Konig H, Lohr M, Edlund K, Botling J, Micke P, Nagase T
    Clin Cancer Res, 2014-06-20;20(17):4660-72.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Autocrine ligands of the epithelial growth factor receptor mediate inflammatory responses to diesel exhaust particles.
    Authors: Parnia S, Hamilton L, Puddicombe S, Holgate S, Frew A, Davies D
    Respir Res, 2014-02-20;15(0):22.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Aberrantly activated AREG-EGFR signaling is required for the growth and survival of CRTC1-MAML2 fusion-positive mucoepidermoid carcinoma cells.
    Authors: Chen, Z, Chen, J, Gu, Y, Hu, C, Li, J-L, Lin, S, Shen, H, Cao, C, Gao, R, Li, J, Ha, P K, Kaye, F J, Griffin, J D, Wu, L
    Oncogene, 2013-08-26;33(29):3869-77.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Activation of HER family signaling as a mechanism of acquired resistance to ALK inhibitors in EML4-ALK-positive non-small cell lung cancer.
    Authors: Tanizaki J, Okamoto I, Okabe T, Sakai K, Tanaka K, Hayashi H, Kaneda H, Takezawa K, Kuwata K, Yamaguchi H, Hatashita E, Nishio K, Nakagawa K
    Clin Cancer Res, 2012-07-27;18(22):6219-26.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. 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
  26. 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
  27. 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
  28. Amphiregulin carboxy-terminal domain is required for autocrine keratinocyte growth.
    Authors: Stoll SW, Johnson JL, Li Y, Rittie L, Elder JT
    J. Invest. Dermatol., 2010-04-29;130(8):2031-40.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. Cross-talk between estrogen receptor and epidermal growth factor receptor in head and neck squamous cell carcinoma.
    Authors: Egloff AM, Rothstein ME, Seethala R, Siegfried JM, Grandis JR, Stabile LP
    Clin. Cancer Res., 2009-10-13;15(21):6529-40.
    Species: Human
    Sample Types: Cell Culture Supernates
  30. 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
  31. Phase II study and biomarker analysis of cetuximab combined with modified FOLFOX6 in advanced gastric cancer.
    Authors: Han SW, Oh DY, Im SA, Park SR, Lee KW, Song HS, Lee NS, Lee KH, Choi IS, Lee MH, Kim MA, Kim WH, Bang YJ, Kim TY
    Br. J. Cancer, 2009-01-06;100(2):298-304.
    Species: Human
    Sample Types: Serum
  32. 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
  33. 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
  34. Epidermal growth factor receptor pathway analysis identifies amphiregulin as a key factor for cisplatin resistance of human breast cancer cells.
    Authors: Eckstein N, Servan K, Girard L, Cai D, von Jonquieres G, Jaehde U, Kassack MU, Gazdar AF, Minna JD, Royer HD
    J. Biol. Chem., 2007-10-17;283(2):739-50.
    Species: Human
    Sample Types: Cell Culture Supernates
  35. The ADAM10 prodomain is a specific inhibitor of ADAM10 proteolytic activity and inhibits cellular shedding events.
    Authors: Moss ML, Bomar M, Liu Q, Sage H, Dempsey P, Lenhart PM, Gillispie PA, Stoeck A, Wildeboer D, Bartsch JW, Palmisano R, Zhou P
    J. Biol. Chem., 2007-09-25;282(49):35712-21.
    Species: Human
    Sample Types: Cell Culture Supernates
  36. 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
  37. Alteration of the serum levels of the epidermal growth factor receptor and its ligands in patients with non-small cell lung cancer and head and neck carcinoma.
    Authors: Lemos-Gonzalez Y, Rodriguez-Berrocal FJ, Cordero OJ, Gomez C, Paez de la Cadena M
    Br. J. Cancer, 2007-04-24;96(10):1569-78.
    Species: Human
    Sample Types: Serum
  38. 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
  39. The response of human epithelial cells to TNF involves an inducible autocrine cascade.
    Authors: Janes KA, Gaudet S, Albeck JG, Nielsen UB, Lauffenburger DA, Sorger PK
    Cell, 2006-03-24;124(6):1225-39.
    Species: Human
    Sample Types: Cell Culture Supernates
  40. 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
  41. 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
  42. Autocrine extracellular signal-regulated kinase (ERK) activation in normal human keratinocytes: metalloproteinase-mediated release of amphiregulin triggers signaling from ErbB1 to ERK.
    Authors: Kansra S, Johnson JL
    Mol. Biol. Cell, 2004-07-14;15(9):4299-309.
    Species: Human
    Sample Types: Cell Culture Supernates


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Human Amphiregulin DuoSet ELISA
By Anonymous on 01/25/2018
Sample Tested: Cell culture supernatant,Lung tissue