Human Phospho-EGFR (Y1086) Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human EGFR when phosphorylated at Y1086 in Western blots.
  • Source
    Monoclonal Rat IgG2A Clone # 938835
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Phosphopeptide containing the human EGFR Y1086 site
    Accession # P00533
  • 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
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    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
Detection of Phospho-EGFR (Y1086) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB89671) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Phospho-EGFR (Y1086) at approximately 180 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
EGFR in A431 Human Cell Line. EGFR phosphorylated at Y1086 was detected in immersion fixed A431 human epithelial carcinoma cell line treated with Recombinant Human EGF (Catalog # 236-EG) using Rat Anti-Human EGFR Monoclonal Antibody (Catalog # MAB89671) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cell membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Phospho-EGFR (Y1086) by Simple WesternTM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 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 (Y1086) at approximately 197 kDa (as indicated) using 10 µg/mL of Rat Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB89671) followed by 1:50 dilution of HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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 Y1086 creates a binding site for the adaptor protein GRB2, 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
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