Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2149Y] to EGFR (phospho Y845)
  • Suitable for: Dot blot, WB, ICC/IF, Flow Cyt
  • Reacts with: Human

Overview

  • Product name

    Anti-EGFR (phospho Y845) antibody [EPR2149Y]
    See all EGFR primary antibodies

  • Description

    Rabbit monoclonal [EPR2149Y] to EGFR (phospho Y845)

  • Host species

    Rabbit

  • Specificity

    ab97613 detects EGFR phosphorylated on tyrosine 845.

  • 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 (phospho S845). The exact sequence is proprietary.

  • Positive control

    • A431 cells and cell lysates.

  • General notes

    Mouse, 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    Western blot - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    All lanes : Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613) at 1/1500 dilution (purified)

    Lane 1 : Untreated A431 (Human epidermoid carcinoma cell line) whole cell lysates
    Lane 2 : A431 (Human epidermoid carcinoma cell line) treated with 100 ng/ml EGF for 10 minutes whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 134 kDa
    Observed band size: 175 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking and dilution buffer: 5% NFDM/TBST. 

  • Flow Cytometry - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    Flow Cytometry - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)

    Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 10 minutes cells labeling EGFR with purified ab97613 at 1/400 dilution (1 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Untreated A431 cells (Green).

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed,0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cell lines labeling EGFR with ab97613 (purified) at 1/50 dilution,followed by Goat anti-Rabbit IgG H & L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution

    Cells were counterstained with ab195889 (Anti-alpha Tubulin antibody [DM1A] ) - Microtubule Marker (Alexa Fluor® 594) 

    The nuclear counterstain is DAPI (blue).

    The green staining on the membrane was increased in the EGF (100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.

    For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment. The data showed mostly membranous staining.

     

  • Dot Blot - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    Dot Blot - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)

    Dot Blot analysis of Lane 1: EGFR (pY845) phospho peptide and Lane 2: EGFR non-phospho peptide labeling EGFR (phospho Y845) with ab97613 (unpurified) at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.

  • Western blot - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    Western blot - Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    All lanes : Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613) at 1/500 dilution (unpurified)

    Lane 1 : A431 cell lysates, untreated
    Lane 2 : A431 cell lysates, treated with EGF

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 134 kDa

  • Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)
    Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613)

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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