Overview

  • Product nameAnti-EGFR antibody [E235]
    See all EGFR primary antibodies
  • Description
    Rabbit monoclonal [E235] to EGFR
  • Specificityab32077 detects Epidermal growth factor receptor (EGFR). It does not cross react with other ERBB family members. 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 applicationsICC/IF, WB, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human EGFR aa 1150 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control
    • WB: A431 cell lysate. ICC/IF: A431 cells. Flow Cyt: A431 cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

    Alternative versions available:

    Anti-EGFR antibody (Alexa Fluor® 647) [E235] (ab205732)

    Anti-EGFR antibody (Alexa Fluor® 488) [E235] (ab205731)

Properties

Applications

Our Abpromise guarantee covers the use of ab32077 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/500.
WB 1/1000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 134 kDa).

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 Cyt 1/10 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/30.
  • Application notesIs unsuitable for IHC.
  • Target

    • Information by UniProt
    • Database links
    • Alternative names
      • Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
      • Cell growth inhibiting protein 40 antibody
      • Cell proliferation inducing protein 61 antibody
      • EGF R antibody
      • EGFR antibody
      • EGFR_HUMAN antibody
      • Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) antibody
      • Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian) antibody
      • Epidermal growth factor receptor antibody
      • erb-b2 receptor tyrosine kinase 1 antibody
      • ERBB antibody
      • ERBB1 antibody
      • Errp antibody
      • HER1 antibody
      • mENA antibody
      • NISBD2 antibody
      • Oncogen ERBB antibody
      • PIG61 antibody
      • Proto-oncogene c-ErbB-1 antibody
      • Receptor tyrosine protein kinase ErbB 1 antibody
      • Receptor tyrosine-protein kinase ErbB-1 antibody
      • SA7 antibody
      • Species antigen 7 antibody
      • Urogastrone antibody
      • v-erb-b Avian erythroblastic leukemia viral oncogen homolog antibody
      • wa2 antibody
      • Wa5 antibody
      see all

    Anti-EGFR antibody [E235] images

    • All lanes : Anti-EGFR antibody [E235] (ab32077) at 1/1000 dilution

      Lane 1 : Caco-2 cell lysate
      Lane 2 : A431 cell lysate
      Lane 3 : Mouse skin cell lysate
      Lane 4 : Rat 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 ab32077 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.

    • Immunocytochemistry/Immunofluorescence analysis of A431 cells labelling EGFR with ab32077 at 1/500. Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
      Control: PBS only.
      Nuclear counter stain: DAPI.

    • Overlay histogram showing A431 cells stained with ab32077 (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 (ab32077, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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.

    References for Anti-EGFR antibody [E235] (ab32077)

    This product has been referenced in:
    • Hofmann K  et al. A novel alkyne cholesterol to trace cellular cholesterol metabolism and localization. J Lipid Res 55:583-91 (2014). Read more (PubMed: 24334219) »

    See 1 Publication for this product

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"