Overview

  • Product nameAnti-EGFR antibody [EP38Y]
    See all EGFR primary antibodies
  • Description
    Rabbit monoclonal [EP38Y] to EGFR
  • SpecificityThe immunogen for this product is a synthetic phospho-peptide corresponding to residues surrounding Tyr1068 of human EGFR. After screening, clone “EP38Y” was found to recognize total EGFR and is not specific to phosphorylated-Tyr1068 EGFR. 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 applicationsWB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human EGFR. Synthetic phospho-peptide corresponding to residues surrounding Tyr1068 of mature human EGFR.
    Database link: P00533

  • Positive control
    • A431, HeLa cell lysate or human lymph node metastasis tissue.
  • General notes

    A trial size is available to purchase for this antibody.

    Alternative versions available:
    Anti-EGFR antibody (Alexa Fluor® 488) [EP38Y] (ab193244)
    Anti-EGFR antibody (Alexa Fluor® 647) [EP38Y] (ab192982)
    Anti-EGFR antibody (HRP) [EP38Y] (ab193602)
    Anti-EGFR antibody (Alexa Fluor® 594) [EP38Y] (ab207870)
    Anti-EGFR antibody (Phycoerythrin) [EP38Y] (ab208753)

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab52894 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
WB 1/1000 - 1/10000. Detects a band of approximately 175 kDa (predicted molecular weight: 134 kDa).Can be blocked with EGFR peptide (ab204282).

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

IP 1/50.
Flow Cyt 1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.

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 [EP38Y] images

  • All lanes : Anti-EGFR antibody [EP38Y] (ab52894) 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 ab52894 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.

  • All lanes : Anti-EGFR antibody [EP38Y] (ab52894) at 1/1000 dilution (Anti-EGFR antibody [EP38Y])

    Lane 1 : H820 siCtl cell lysate, siRNA treated
    Lane 2 : H820 siEGFR#1 cell lysate, siRNA treated
    Lane 3 : H820 siEGFR#2 cell lysate, siRNA treated

    Lysates/proteins at 20 µg per lane.

    Secondary
    GE Healthcare Polyclonal Anti Rabbit, HRP conjugated at 1/5000 dilution

    Predicted band size : 134 kDa
    Observed band size : 165 kDa (why is the actual band size different from the predicted?)

    Gel running conditions: 4-15%, reduced and denatured.

    Blocking agent: 5% milk.

    Blocking time: 1 hour.

    Incubation (with primary antibody): 16 hours, 4 Celsius.

    Dilution buffer: 5%BSA

  • ab52894 showing positive staining in Endometrial carcinoma tissue.

  • Overlay histogram showing A431 cells stained with ab52894 (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 (ab52894, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG H+L) (ab96899) at 1/500 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.

  • Anti-EGFR antibody [EP38Y] (ab52894) at 1/100000 dilution + HeLa cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labeled. at 1/2000 dilution

    Predicted band size : 134 kDa
    Observed band size : 175 kDa (why is the actual band size different from the predicted?)
  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Ab52894 (1:100) staining human EGFR in human lymph node metastasis tissue by immunohistochemistry using paraffin embedded tissue.Ab52894 (1:100) staining human EGFR in human lymph node metastasis tissue by immunohistochemistry using paraffin embedded tissue.
  • Anti-EGFR antibody [EP38Y] (ab52894) at 1/500 dilution + Murine macrophage lysates at 80 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 134 kDa
    Additional bands at : 170 kDa (possible post-translational modification).

    Exposure time : 1 minute

    Image is courtesy of an anonymous AbReview.

    Blocking was with 5% BSA incubated for 1 hour at 25°C.

    See Abreview

  • ab52894 showing positive staining in Cervical carcinoma tissue.

  • ab52894 showing positive staining in Glioma tissue.

  • ab52894 showing positive staining in Normal tonsil squamous cells tissue.

  • ab52894 showing positive staining in Renal cell carcinoma tissue.

References for Anti-EGFR antibody [EP38Y] (ab52894)

This product has been referenced in:
  • Shuai S  et al. TIP30 nuclear translocation negatively regulates EGF-dependent cyclin D1 transcription in human lung adenocarcinoma. Cancer Lett 354:200-9 (2014). Read more (PubMed: 25135222) »
  • Samkoe KS  et al. Quantitative in vivo immunohistochemistry of epidermal growth factor receptor using a receptor concentration imaging approach. Cancer Res 74:7465-74 (2014). Read more (PubMed: 25344226) »

See all 12 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (Human Renal Epithelial Cells)
Gel Running Conditions Non-reduced Non-Denaturing (Native) (8%)
Loading amount 60 µg
Treatment 20ng EGF for 30min
Specification Human Renal Epithelial Cells
Blocking step BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. 腾飞 白

Verified customer

Submitted Jan 06 2016

Application Western blot
Sample Mouse Cell lysate - other (macrophage)
Gel Running Conditions Reduced Denaturing
Loading amount 80 µg
Specification macrophage
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 06 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (D3 mouse embryonic stem cells)
Permeabilization Yes - Triton 0.1%
Specification D3 mouse embryonic stem cells
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1%
Fixative Paraformaldehyde
Username

Miss. Fabiana Luise

Verified customer

Submitted Oct 22 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (4-15%)
Sample Human Cell lysate - whole cell (non small cell lung cancer)
Specification non small cell lung cancer
Treatment siRNA treated
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 07 2014


The pretreatment note in the protocol of the IHC kit refers to the antigen retrieval step that is required for most antibody to achieve a successful IHC-P staining.
For both of these antibodies (ab14440 and ab52894) I can recommend to use Sod...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton X100 in PBS
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 25 2014

Hello! We do recommend you use an antigen retrieval step, heat mediated such as citrate pH6. The antibody you suggest, ab52894 is a good one. Another EGFR antibody with good feedback is ab9482. We do guarantee our products to work as specified on the d...

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Thank you for contacting us.

Rabbit monoclonals typically have 10-100x the affinity of mouse monoclonals, as such it is often the case that the concentration of a rabbit monoclonal will be lower than a mouse monoclonal. Ab52894 does not ha...

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Paraffin blocks are commonly cut into 4 µm thickness sections and those sections are stored in a sealed box at room temperature for no more than 6 months. We have not observed any particular rapid degradation of the EGFR antigen.

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