Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-BNIP3 antibody [EPR4034] (ab109362)

Reviews (3) Submit a question References (27)

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

    Suitable for: WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human BNIP3 aa 100-200. The exact sequence is proprietary.

  • 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

Applications

Our Abpromise guarantee covers the use of ab109362 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. Detects a band of approximately 30 kDa (predicted molecular weight: 22 kDa).

For unpurified use at 1/1000 - 1/10000.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

(Heat to 98°C, allow to cool for 10-20 minutes).

For unpurified use at 1/50 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/150.

For unpurified use at 1/100 - 1/250.

Target

  • Function

    Apoptosis-inducing protein that, which can overcome BCL2 suppression. May play a role in repartitioning calcium between the two major intracellular calcium stores in association with BCL2.

  • Sequence similarities

    Belongs to the NIP3 family.

  • Cellular localization

    Mitochondrion. Mitochondrion membrane. Coexpression with the EIB 19-kDa protein results in a shift in NIP3 localization pattern to the nuclear envelope. Colocalizes with ACAA2 in the mitochondria.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • BCL2 Adenovirus E1B 19kDa Interacting Protein 3 antibody
    • BCL2/adenovirus E1B 19 kDa protein interacting protein 3 antibody
    • BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 antibody
    • BNIP 3 antibody
    • BNIP3 antibody
    • BNIP3_HUMAN antibody
    • NIP 3 antibody
    • NIP3 antibody
    see all

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.

References

Publishing research using ab109362? Please let us know so that we can cite the reference in this datasheet.

ab109362 has been referenced in 27 publications.

  • Zhang J  et al. Involvement of autophagy in hypoxia-BNIP3 signaling to promote epidermal keratinocyte migration. Cell Death Dis 10:234 (2019). PubMed: 30850584
  • Xie M  et al. Trimetazidine combined with exercise improves exercise capacity and anti-fatal stress ability through enhancing mitochondrial quality control. Life Sci 224:157-168 (2019). PubMed: 30872179
  • Cai CC  et al. Glycine Protects against Hypoxic-Ischemic Brain Injury by Regulating Mitochondria-Mediated Autophagy via the AMPK Pathway. Oxid Med Cell Longev 2019:4248529 (2019). PubMed: 30881590
  • Niu Y  et al. RNA N6-methyladenosine demethylase FTO promotes breast tumor progression through inhibiting BNIP3. Mol Cancer 18:46 (2019). PubMed: 30922314
  • Pietzsch S  et al. Modulation of cardiac AKT and STAT3 signalling in preclinical cancer models and their impact on the heart. Biochim Biophys Acta Mol Cell Res N/A:118519 (2019). PubMed: 31374232
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-BNIP3 antibody [EPR4034]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver and Kidney lysates)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
30 µg
Specification
Liver and Kidney lysates
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Dec 27 2019

Western blot abreview for Anti-BNIP3 antibody [EPR4034]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Pig Tissue lysate - whole (Heart)
Gel Running Conditions
Non-reduced Denaturing (4-15%)
Loading amount
20 µg
Specification
Heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Read More

Antoine Chaanine

Verified customer

Submitted Mar 31 2017

Western blot abreview for Anti-BNIP3 antibody [EPR4034]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (mouse lung)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
mouse lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted Feb 20 2017

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