Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-BNIP3 antibody [EPR4034] (ab109362)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4034] to BNIP3
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-BNIP3 antibody [EPR4034]
    See all BNIP3 primary antibodies

  • Description

    Rabbit monoclonal [EPR4034] to BNIP3

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Rat
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Jurkat cells, Jurkat cells treated with etoposide, SH-SY5Y cells, SH-SY5Y cells treated with camptothecin and MCF-7 cells treated with Cocl2 lysates and rat kidney, mouse spleen and mouse kidney tissue lysates. IHC-P: Human renal adenocarcinoma and kidney tissues. ICC/IF: HeLa and SH-SY5Y cells.

  • General notes

     

     

    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-BNIP3 antibody [EPR4034] (ab109362)
    Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)

    Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
    Lane 2: BNIP3 knockout HAP1 whole cell lysate (40 µg)
    Lane 3: SHSY5Y whole cell lysate (40 µg)
    Lane 4: A431 whole cell lysate (40 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109362 observed at 25 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab109362 was shown to recognize BNIP3 in wild-type HAP1 cells as signal was lost at the expected MW in BNIP3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and BNIP3 knockout samples were subjected to SDS-PAGE. ab109362 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Anti-BNIP3 antibody [EPR4034] (ab109362) at 1/1000 dilution (purified) + SH-SY5Y cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Anti-BNIP3 antibody [EPR4034] (ab109362) at 1/2000 dilution (purified) + Jurkat cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 30 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    All lanes : Anti-BNIP3 antibody [EPR4034] (ab109362) at 1/2000 dilution (purified)

    Lane 1 : Rat kidney tissue lysate
    Lane 2 : Mouse kidney tissue lysate
    Lane 3 : Mouse spleen tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 30 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362)
    All lanes : Anti-BNIP3 antibody [EPR4034] (ab109362) at 1/1000 dilution (unpurified)

    Lane 1 : Jurkat cell lysate treated with etoposide
    Lane 2 : Jurkat cell lysate
    Lane 3 : SH-SY5Y cell lysate treated with camptothecin
    Lane 4 : SH-SY5Y cell lysate
    Lane 5 : MCF-7 cell lysate treated with Cocl2

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 22 kDa
    Observed band size: 30 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (ab109362)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human renal adenocarcinoma tissue labelling BNIP3 with purified ab109362 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (ab109362)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling BNIP3 with unpurified ab109362 at a 1/50 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-BNIP3 antibody [EPR4034] (ab109362)
    Immunocytochemistry/ Immunofluorescence - Anti-BNIP3 antibody [EPR4034] (ab109362)

    Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling BNIP3 with unpurified ab109362 at a 1/100 dilution.

  • Anti-BNIP3 antibody [EPR4034] (ab109362)
    Anti-BNIP3 antibody [EPR4034] (ab109362)

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