Human Phospho-EGFR (Y1173) Antibody

(5 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human EGFR when phosphorylated at Y1173 in Western blots.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Phosphopeptide containing human EGFR Y1173 site
  • 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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Simple Western
    2 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
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
Intracellular Staining by Flow Cytometry
Detection of Phospho-EGFR (Y1173) in A431 Human Cell Line by Flow Cytometry. A431 human epithelial carcinoma cells were untreated (open histogram), or treated for 5 minutes with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1095), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with para­formaldehyde and permeabilized with saponin.
Detection of Human Phospho-EGFR (Y1173) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated
(‑) or treated (+) with 100 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGFR (Y1173) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Phospho-EGFR (Y1173) in A431 Human Cell Line. EGFR phosphorylated at Y1173 was detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
Phospho-EGFR (Y1173) in Mouse Embryo. EGFR phosphorylated at Y1173 was detected in immersion fixed frozen sections of mouse embryo using Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) 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 Frozen Tissue Sections.
Detection of Human Phospho-EGFR (Y1173) by Simple WesternTM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-EGFR (Y1173) at approximately 265 kDa (as indicated) using 2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.
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: EGFR
Epidermal growth factor receptor (EGFR, also known as ErbB1 and HER1) is the founding member of the ErbB family of receptor tyrosine kinases. Ligand binding induces receptor dimerization and autophosphorylation on multiple tyrosine residues. Phosphorylation of Y1173 creates a binding site for the protein tyrosine phosphatase SHP-1, leading to attenuation of receptor signaling.
  • Long Name:
    Epidermal Growth Factor Receptor
  • Entrez Gene IDs:
    1956 (Human); 13649 (Mouse)
  • Alternate Names:
    avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; EC 2.7.10; EC 2.7.10.1; EGF R; EGFR; epidermal growth factor receptor (avian erythroblastic leukemia viral (v-erb-b)oncogene homolog); epidermal growth factor receptor; ErbB; ErbB1; ERBB1PIG61; HER1; HER-1; mENA; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1
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.

5 Citations: Showing 1 - 5
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Species
Applications
Sample Type
  1. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016;1(13):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC-Fluorescent
  2. Phenotypic diversity of breast cancer-related mutations in metalloproteinase-disintegrin ADAM12.
    Authors: Qi, Yue, Duhachek-Muggy, Sara, Li, Hui, Zolkiewska, Anna
    PLoS ONE, 2014;9(3):e92536.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: WB
  3. Analysis of Epithelial Growth Factor-Receptor (EGFR) Phosphorylation in Uterine Smooth Muscle Tumors: Correlation to Mucin-1 and Galectin-3 Expression.
    Authors: Weissenbacher, Tobias, Vrekoussis, Thomas, Roeder, David, Makrigiannakis, Antonis, Mayr, Doris, Ditsch, Nina, Friese, Klaus, Jeschke, Udo, Dian, Darius
    Int J Mol Sci, 2013;14(3):4783-92.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  4. Repulsion of cerebellar granule neurons by chondroitin sulfate proteoglycans is mediated by MAPK pathway.
    Authors: Kaneko M, Kubo T, Hata K, Yamaguchi A, Yamashita T
    Neurosci. Lett., 2007;423(1):62-7.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  5. Protein C is an autocrine growth factor for human skin keratinocytes.
    Authors: Xue M, Campbell D, Jackson CJ
    J. Biol. Chem., 2007;282(18):13610-6.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
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