Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5562] to EBP50/NHERF-1
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-EBP50/NHERF-1 antibody [EPR5562]
    See all EBP50/NHERF-1 primary antibodies

  • Description

    Rabbit monoclonal [EPR5562] to EBP50/NHERF-1

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • HCT116, HepG2, 293T, Jurkat, C6, PC-12 and MCF-7 cell lysates; Human kidney tissue

  • General notes

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with this 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-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
    Western blot - Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
    All lanes : Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : SLC9A3R1 knockout HCT116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 39 kDa
    Observed band size: 48 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab109430 observed at 48 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab109430 was shown to react with EBP50/NHERF-1 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266876 (knockout cell lysate ab257281) was used. Wild-type HCT116 and SLC9A3R1 knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109430 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)

    Immunohistochemical analysis of EBP50/NHERF-1 in paraffin-embedded Human kidney tissue using ab109430 at 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
    Western blot - Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: EBP50/NHERF-1 knockout HAP1 cell lysate (20 µg)
    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab109430 observed at 48 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab109430 was shown to specifically recognize EBP50/NHERF-1 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when EBP50/NHERF-1 knockout samples were usexamined. Wild-type and EBP50 knockout samples were subjected to SDS-PAGE. ab109430 and ab18058 (loading control to Vinculin) were diluted at 1/500 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
    Western blot - Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
    All lanes : Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430) at 1/1000 dilution

    Lane 1 : HepG2 cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : C6 cell lysate
    Lane 5 : PC12 cell lysate
    Lane 6 : MCF7 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 39 kDa

  • Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
    Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)

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