Human Amphiregulin Antibody

(12 citations)   
  • 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 as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
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.
Data Examples
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 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 Poly­clonal 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 counter­stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
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.
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.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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
Related Research Areas
Citations:

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.

12 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. 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;20(17):4660-72.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Not Specified
  2. 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, 2014;33(29):3869-77.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Not Specified
  3. 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;288(15):10338-48.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. 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;17(15):5197-204.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  5. 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;15(21):6529-40.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  6. 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;121(0):3608-18.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  7. 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;283(2):739-50.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  8. 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;30(6):1064-73.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  9. 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;77(1):53-60.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  10. 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;37(5):578-88.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  11. 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;133(4):751-9.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  12. 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;15(9):4299-309.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
Expand to show all 12 Citations
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