Human EGFR (aa 746-750 deletion) Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human EGFR with the aa 746-750 deletion in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG2A Clone # 752502
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Human EGFR synthetic peptide
    CPVAIKTSPKAN
    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
    2 µg/mL
    See below
  • Simple Western
    20 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 Human EGFR (aa 746-750 deletion) by Western Blot. Western blot shows lysates of HCC827 human non-small cell lung cancer cell line, HCT‑116 human colorectal carcinoma cell line, and A431 human epithelial carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human EGFR (aa 746-750 deletion) Monoclonal Antibody (Catalog # MAB8336) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for EGFR (aa 746-750 deletion) at approximately 170 kDa (as indicated). For additional reference total EGFR was detected using Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # AF231). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
EGFR (aa 746-750 deletion) in HCC827 Human Cell Line. EGFR (aa 746-750 deletion) was detected in immersion fixed HCC827 human non-small cell lung cancer cell line using Mouse Anti-Human EGFR (aa 746-750 deletion) Monoclonal Antibody (Catalog # MAB8336) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. Negative staining shown in the lower panel with NCI-H1975 human lung epithelial cell line. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human EGFR by Simple WesternTM. Simple Western lane view shows lysates of HCC827 human non-small cell lung cancer cell line, loaded at 0.5 mg/mL. A specific band was detected for EGFR at approximately 200 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human EGFR (aa 746-750 deletion) Monoclonal Antibody (Catalog # MAB8336). This experiment was conducted under reducing conditions and using the
66-440 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. EGFR exon 19 deletions are in-frame deletions occurring within exon 19, which encodes part of the kinase domain. This mutation occurs with a frequency of approximately 48% in EGFR mutant lung tumors. It affects the catalytic domain (amino acids 746-750), and is predominantly associated with non-small cell lung cancer (1). In a metastatic setting, EGFR deletions like aa746-750 are predictors of efficacy of the EGFR tyrosine kinase inhibitors (1, 2).

  • References:
    1. Lynch, T. et al. (2004) N Engl J Med. 350:2129.
    2. Carey, K. et al. (2006) Cancer Res 66:8163.
  • 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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