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Human Amphiregulin Antibody

Catalog #: AF262
35 Citations Datasheet
Catalog # Availability Size / Price Qty
AF262
AF262-SP
Detection of Human Amphiregulin by Western Blot.
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Citations (35)
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Human Amphiregulin Antibody Summary

Species Reactivity
Human
Specificity
Detects human Amphiregulin in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant mouse (rm) Amphiregulin, rmEpiregulin, recombinant human (rh) HB-EGF, and rhTGF-alpha is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Amphiregulin
Val107-Lys184
Accession # P15514
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Immunocytochemistry
5-15 µg/mL
See below
Neutralization
Measured by its ability to neutralize Amphiregulin-induced proliferation in the Balb/3T3 mouse embryonic fibroblast cell line. The Neutralization Dose (ND50) is typically 0.3-1 µg/mL in the presence of 50 ng/mL Recombinant Human Amphiregulin.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Amphiregulin antibody by Western Blot. View Larger

Detection of Human Amphiregulin by Western Blot. Western blot shows conditioned media from T47D human breast cancer cell line untreated (-) or treated (+) with PMA and PHA for 3 days. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Amphiregulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF262) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Amphiregulin at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry Amphiregulin antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC). View Larger

Amphiregulin in MCF‑7 Human Cell Line. Amphiregulin was detected in immersion fixed MCF-7 human breast cancer cell line using Goat Anti-Human Amphiregulin Antigen Affinity-purified Poly-clonal Antibody (Catalog # AF262) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry Amphiregulin antibody in Human Breast by Immunohistochemistry (IHC-P). View Larger

Amphiregulin in Human Breast. Amphiregulin was detected in immersion fixed paraffin-embedded sections of human breast using Goat Anti-Human Amphiregulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF262) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Neutralization Cell Proliferation Induced by Amphiregulin and Neutralization by Human Amphiregulin Antibody. View Larger

Cell Proliferation Induced by Amphiregulin and Neutralization by Human Amphiregulin Antibody. Recombinant Human Amphiregulin (Catalog # 262-AR) stimulates proliferation in the Balb/3T3 mouse embryonic fibroblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Amphiregulin (50 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Amphiregulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF262). The ND50 is typically 0.3-1 µg/mL.

Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Amphiregulin

Amphiregulin (AR) is a member of the EGF family of cytokines which is comprised of at least ten proteins including EGF, TGF-alpha, HB-EGF, and the various heregulins. All of these cytokines are synthesized as transmembrane precursors and are characterized by the presence of one or several EGF structural units in their extracellular domain. The soluble forms of these cytokines are released by proteolytic cleavage. Amphiregulin was originally isolated from the conditioned media of a PMA-treated MCF-7 human breast carcinoma cell line. The AR cDNA encodes a 252 amino acid (aa) residue transmembrane precursor. Multiple forms of native AR containing either 78 or 84 aa residues and both N- and O-linked oligosaccharides have been found. Amphiregulin mRNA expression can be detected in numerous carcinoma cell lines and the epithelial cells of various human tissues including colon, stomach, breast, ovary, kidney, etc.

Human AR stimulates the proliferation of various human and mouse keratinocytes, mammary epithelial cells and some fibroblasts. AR is also a growth inhibitor for various tumor cell lines. In certain colon carcinoma cell lines, AR has been shown to be an autocrine growth factor. Amphiregulin can bind to the EGF receptor. It has been suggested that in certain cell types, AR bioactivity may be mediated through the EGF receptor. The 98 aa residue long form of recombinant amphiregulin has shown to be approximately 5 - 10 fold more active than the 78 aa residue form of recombinant AR in an in vitro proliferation assay using Balb/3T3 fibroblasts.

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

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Citations for Human Amphiregulin Antibody

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.

35 Citations: Showing 1 - 10
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  1. Cell surface annexins regulate ADAM-mediated ectodomain shedding of proamphiregulin.
    Authors: Nakayama H, Fukuda S, Inoue H et al.
    Mol Biol Cell
  2. CITED1 as a marker of favourable outcome in anti-endocrine treated, estrogen-receptor positive, lymph-node negative breast cancer
    Authors: Dahlgren, M;Lettiero, B;Dalal, H;Mårtensson, K;Gaber, A;Nodin, B;Gruvberger-Saal, SK;Saal, LH;Howlin, J;
    BMC research notes
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Accessory ESCRT‐III proteins are conserved and selective regulators of Rab11a‐exosome formation
    Authors: Pauline P. Marie, Shih‐Jung Fan, John Mason, Adam Wells, Cláudia C. Mendes, S. Mark Wainwright et al.
    Journal of Extracellular Vesicles
  4. Luteinizing hormone stimulates the expression of amphiregulin in human theca cells
    Authors: Y Liu, Y Zhong, X Shen, X Guo, R Wu, T Yang, M Chen
    Journal of ovarian research, 2022-12-07;15(1):129.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  5. Mitochondrial stress induces AREG expression and epigenomic remodeling through c-JUN and YAP-mediated enhancer activation
    Authors: Y Hino, K Nagaoka, S Oki, K Etoh, S Hino, M Nakao
    Nucleic Acids Research, 2022-09-23;50(17):9765-9779.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  6. Pan-cancer distribution of cleaved cell-surface amphiregulin, the target of the GMF-1A3 antibody drug conjugate
    Authors: Kristopher A Lofgren, Nicolette C Reker, Sreeja Sreekumar, Paraic A Kenny
    Antibody Therapeutics
  7. Tissue remodeling by an opportunistic pathogen triggers allergic inflammation
    Authors: Karen Agaronyan, Lokesh Sharma, Bharat Vaidyanathan, Keith Glenn, Shuang Yu, Charles Annicelli et al.
    Immunity
  8. AXL cooperates with EGFR to mediate neutrophil elastase-induced migration of prostate cancer cells
    Authors: Zhiguang Xiao, Stephen R. Hammes
    iScience
  9. Anti-tumor efficacy of an MMAE-conjugated antibody targeting cell surface TACE/ADAM17-cleaved Amphiregulin in breast cancer
    Authors: Kristopher A Lofgren, Sreeja Sreekumar, E Charles Jenkins Jr, Kyle J Ernzen, Paraic A Kenny
    Antibody Therapeutics
  10. Glutamine deprivation alters the origin and function of cancer cell exosomes
    Authors: Shih‐Jung Fan, Benjamin Kroeger, Pauline P Marie, Esther M Bridges, John D Mason, Kristie McCormick et al.
    The EMBO Journal
  11. Proximal Tubule–Derived Amphiregulin Amplifies and Integrates Profibrotic EGF Receptor Signals in Kidney Fibrosis
    Authors: Eirini Kefaloyianni, Manikanda Raja Keerthi Raja, Julian Schumacher, Muthu Lakshmi Muthu, Vaishali Krishnadoss, Sushrut S. Waikar et al.
    Journal of the American Society of Nephrology
  12. Locally advanced rectal cancer transcriptomic-based secretome analysis reveals novel biomarkers useful to identify patients according to neoadjuvant chemoradiotherapy response
    Authors: LMD Canto, SS Cury, MC Barros-Fil, BEC Kupper, MDFS Begnami, C Scapulatem, RF Carvalho, FA Marchi, DA Olsen, JS Madsen, BM Havelund, S Aguiar, SR Rogatto
    Sci Rep, 2019-06-18;9(1):8702.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  13. ADAM17-dependent signaling is required for oncogenic human papillomavirus entry platform assembly
    Authors: Snježana Mikuličić, Jérôme Finke, Fatima Boukhallouk, Elena Wüstenhagen, Dominik Sons, Yahya Homsi et al.
    eLife
  14. Amphiregulin Regulates Phagocytosis-Induced Cell Death in Monocytes via EGFR and Matrix Metalloproteinases
    Authors: C Platen, S Dreschers, LK Reiss, J Wappler, TW Orlikowsky
    Mediators Inflamm., 2018-11-04;2018(0):4310419.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  15. Yes-associated protein promotes the abnormal proliferation of psoriatic keratinocytes via an amphiregulin dependent pathway
    Authors: J Jia, C Li, J Yang, X Wang, R Li, S Luo, Z Li, J Liu, Z Liu, Y Zheng
    Sci Rep, 2018-10-15;8(1):14513.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  16. Deoxycholic acid activates epidermal growth factor receptor and promotes intestinal carcinogenesis by ADAM17-dependent ligand release
    Authors: W Dong, L Liu, Y Dou, M Xu, T Liu, S Wang, Y Zhang, B Deng, B Wang, H Cao
    J. Cell. Mol. Med., 2018-06-29;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  17. Combinations of Bevacizumab and Erlotinib show activity in colorectal cancer independent of RAS status
    Authors: P Mésange, A Bouygues, N Ferrand, M Sabbah, AE Escargueil, A Savina, B Chibaudel, C Tournigand, T Andre, A de Gramont, AK Larsen
    Clin. Cancer Res., 2018-02-28;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: IHC
  18. Abnormal amphiregulin expression correlates with gastric cancer prognosis
    Authors: Bing Wang, Hongmei Yong, Huijun Zhu, Daguang Ni, Sijie Tang, Shu Zhang et al.
    Oncotarget
  19. Epidermal growth factor receptor signalling regulates granulocyte-macrophage colony-stimulating factor production by airway epithelial cells and established allergic airway disease
    Authors: Thomas H. Acciani, Takuji Suzuki, Bruce C. Trapnell, Timothy D. Le Le Cras
    Clinical & Experimental Allergy
  20. Amphiregulin Is a Critical Downstream Effector of Estrogen Signaling in ER alpha -Positive Breast Cancer
    Authors: Esther A. Peterson, Edmund C. Jenkins, Kristopher A. Lofgren, Natasha Chandiramani, Hui Liu, Evelyn Aranda et al.
    Cancer Research
  21. Proteins of the VEGFR and EGFR pathway as predictive markers for adjuvant treatment in patients with stage II/III colorectal cancer: results of the FOGT-4 trial
    Authors: Thomas Thomaidis, Annett Maderer, Andrea Formentini, Susanne Bauer, Mario Trautmann, Michael Schwarz et al.
    Journal of Experimental & Clinical Cancer Research
  22. 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: Whole Tissue
    Applications: IHC
  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: Whole Tissue
    Applications: IHC
  24. Human antigen R-mediated mRNA stabilization is required for ultraviolet B-induced autoinduction of amphiregulin in keratinocytes.
    Authors: Nakayama H, Fukuda S, Matsushita N, Nishida-Fukuda H, Inoue H, Shirakata Y, Hashimoto K, Higashiyama S
    J Biol Chem, 2013-02-21;288(15):10338-48.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  25. Regulation of Amphiregulin Gene Expression by beta -Catenin Signaling in Human Hepatocellular Carcinoma Cells: A Novel Crosstalk between FGF19 and the EGFR System
    Authors: Maria U. Latasa, Fabiana Salis, Raquel Urtasun, Oihane Garcia-Irigoyen, Maria Elizalde, Iker Uriarte et al.
    PLoS ONE
  26. Knock-down of amphiregulin inhibits cellular invasion in inflammatory breast cancer
    Authors: Andrea Baillo, Craig Giroux, Stephen P. Ethier
    Journal of Cellular Physiology
  27. Expression of amphiregulin and EGFRvIII affect outcome of patients with squamous cell carcinoma of the head and neck receiving cetuximab-docetaxel treatment.
    Authors: Tinhofer I, Klinghammer K, Weichert W, Knodler M, Stenzinger A, Gauler T, Budach V, Keilholz U
    Clin. Cancer Res., 2011-06-08;17(15):5197-204.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  28. 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: Whole Cells
    Applications: Functional Assay
  29. Plasma-membrane-anchored growth factor pro-amphiregulin binds A-type lamin and regulates global transcription.
    Authors: Isokane M, Hieda M, Hirakawa S, Shudou M, Nakashiro K, Hashimoto K, Hamakawa H, Higashiyama S
    J. Cell. Sci., 2008-11-01;121(0):3608-18.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  30. 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: Whole Cells
    Applications: Neutralization
  31. 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: Whole Cells
    Applications: Neutralization
  32. EGFR signaling is required for TGF-beta 1 mediated COX-2 induction in human bronchial epithelial cells.
    Authors: Liu M, Yang SC, Sharma S, Luo J, Cui X, Peebles KA, Huang M, Sato M, Ramirez RD, Shay JW, Minna JD, Dubinett SM
    Am. J. Respir. Cell Mol. Biol., 2007-06-28;37(5):578-88.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  33. Epidermal growth factor-like growth factors prevent apoptosis of alcohol-exposed human placental cytotrophoblast cells.
    Authors: Wolff GS, Chiang PJ, Smith SM, Romero R, Armant DR
    Biol. Reprod., 2007-03-28;77(1):53-60.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  34. Human trophoblast survival at low oxygen concentrations requires metalloproteinase-mediated shedding of heparin-binding EGF-like growth factor.
    Authors: Armant DR, Kilburn BA, Petkova A, Edwin SS, Duniec-Dmuchowski ZM, Edwards HJ, Romero R, Leach RE
    Development, 2006-01-11;133(4):751-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  35. 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: Whole Cells
    Applications: Neutralization

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