Human Phospho-EGFR (Y1086) Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human EGFR when phosphorylated at Y1086 in Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1240C
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Phosphopeptide containing the human EGFR  Y1086 site
    Accession # P00533
  • Formulation
    Supplied as a solution in PBS containing BSA, Glycerol and Sodium Azide. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1:1000 dilution
    See below
  • Simple Western
    1:100 dilution
    See below
  • Immunocytochemistry
    1:300-1:500 dilution
    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 Human Phospho-EGFR (Y1086) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line and HeLa human cervical 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 1:1000 dilution of Rabbit Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB8967) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). 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
Phospho-EGFR (Y1086) in A431 Human Cell Line. EGFRphosphorylated at Y1086 was detected in immersion fixed A431 human epithelial carcinoma cell line treated with Recombinant Human EGF (Catalog # 236‑EG) using Rabbit Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB8967) at a 1:300 dilution 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). 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 199 kDa (as indicated) using 1:100 dilution of Rabbit Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB8967). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. 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 opening.
    • 6 months, -20 to -70 °C under sterile conditions after opening.
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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