Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y39] to EGFR (phospho Y1086) - BSA and Azide free
  • Suitable for: Dot blot, ICC/IF, WB, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free
    See all EGFR primary antibodies

  • Description

    Rabbit monoclonal [Y39] to EGFR (phospho Y1086) - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody only detects EGFR phosphorylated on Tyrosine 1086 of the mature human isoform 1.

  • Tested applications

    Suitable for: Dot blot, ICC/IF, WB, IPmore details
    Unsuitable for: Flow Cyt or IHC

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IP: A431 treated with 100ng/mL EGF for 30min whole cell lysate.

  • General notes

    ab247225 is the carrier-free version of ab32086.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

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

Images

  • Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/3000 dilution

    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: 170 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    This data was developed using ab32086, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    Immunocytochemistry - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    This data was developed using ab32086, the same antibody clone in a different buffer formulation.Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR (phospho Y1086) with ab32086 at 1/100, 3 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, a AlexaFluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000, 2 μg/ml. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclear stain was 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.
  • Immunoprecipitation - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    Immunoprecipitation - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)

    This data was developed using ab32086, the same antibody clone in a different buffer formulation.
    Purified ab32086 at 1/20 dilution (1µg) immunoprecipitating EGFR in A431 treated with 100ng/mL EGF for 30min whole cell lysate.
    Lane 1 (input): A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/mL EGF for 30min whole cell lysate 10µg
    Lane 2 (+): ab32086 + A431 treated with 100ng/mL EGF for 30min whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32086 in A431 treated with 100ng/mL EGF for 30min whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 175 kDa

  • Dot Blot - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    Dot Blot - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    This data was developed using ab32086, the same antibody clone in a different buffer formulation.Dot blot analysis of EGFR (phospho Y1086) phospho peptide (Lane 1), EGFR non-phospho peptide (Lane 2), labeling EGFR (phospho Y1086) with ab32068 at a dilution of 1/1000. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000. Blocking and dilution buffer: 5% NFDM/TBST. Exposure time: 5 seconds.
  • Immunocytochemistry - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    Immunocytochemistry - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    This data was developed using ab32086, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of EGFR (phospho Y1086) expression in A431 cell culture, using 1/100 ab32086.
  • Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/10000 dilution

    Lane 1 : Untreated A431 cell lysate
    Lane 2 : A431 cell lysate, treated with EGF

    Predicted band size: 134 kDa
    Observed band size: 150 kDa why is the actual band size different from the predicted?



    This data was developed using ab32086, the same antibody clone in a different buffer formulation.

  • Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)
    Anti-EGFR (phospho Y1086) antibody [Y39] - BSA and Azide free (ab247225)

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