Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-EGFR antibody [EPR39Y] (ab76153)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR39Y] to EGFR
  • Suitable for: ICC/IF, WB, Flow Cyt
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-EGFR antibody [EPR39Y]
    See all EGFR primary antibodies

  • Description

    Rabbit monoclonal [EPR39Y] to EGFR

  • Host species

    Rabbit

  • Specificity

    ab76153 detects both phosphorylated and non-phosphorylated EGFR protein. This product yielded a strong signal in western blot using A431 (human squamous carcinoma) lysate which naturally overexpresses the EGFR protein. Western blot conditions may need to be optimised for cell lines and tissues that express lower levels of endogenous EGFR.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human EGFR. The exact sequence is proprietary. A synthetic peptide corresponding to residues surrounding Tyr1068 of mature human EGFR
    Database link: P00533

  • Positive control

    • HeLa whole cell lysate (ab150035); A431 cells; Ms skin tissue lysate.

  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-EGFR antibody [EPR39Y] (ab76153)
    Western blot - Anti-EGFR antibody [EPR39Y] (ab76153)
    All lanes : Anti-EGFR antibody [EPR39Y] (ab76153) at 1/1000 dilution

    Lane 1 : Caco-2 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Mouse skin cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 134 kDa



    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab76153 overnight at 4°C in the presence of loading control ab18058 (Mouse monoclonal [SPM227] to Vinculin diluted 1:10000). Antibody binding was detected using IR-labelled goat anti-Rabbit Ab at a 1:10,000 dilution for one hour at room temperature before imaging.

    This product yielded a strong signal in western blot using A431 (human squamous carcinoma) lysate which naturally overexpresses the EGFR protein.  Western blot conditions may need to be optimised for cell lines and tissues that express lower levels of endogenous EGFR.

  • Flow Cytometry - Anti-EGFR antibody [EPR39Y] (ab76153)
    Flow Cytometry - Anti-EGFR antibody [EPR39Y] (ab76153)
    Overlay histogram showing A431 cells stained with ab76153 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76153, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EPR39Y] (ab76153)
    Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EPR39Y] (ab76153)
    ab76153 at 1/100 dilution staining EGFR in A431 cells by Immunofluorescence.
  • Western blot - Anti-EGFR antibody [EPR39Y] (ab76153)
    Western blot - Anti-EGFR antibody [EPR39Y] (ab76153)
    Anti-EGFR antibody [EPR39Y] (ab76153) at 1/5000 dilution + HeLa cell lysate at 10 µg

    Secondary
    goat anti-rabbit HRP at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 134 kDa
    Observed band size: 170,180 kDa
    why is the actual band size different from the predicted?

  • Anti-EGFR antibody [EPR39Y] (ab76153)
    Anti-EGFR antibody [EPR39Y] (ab76153)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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