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Anti-BNIP3 antibody [EPR4034] ab109362 is a rabbit monoclonal antibody that is used in BNIP3 western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with BNIP3 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR4034 is cited in over 80 publications


Images

Western blot - Anti-BNIP3 antibody [EPR4034] (AB109362), expandable thumbnail
  • Western blot - Anti-BNIP3 antibody [EPR4034] (AB109362), expandable thumbnail
  • Western blot - Anti-BNIP3 antibody [EPR4034] (AB109362), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (AB109362), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (AB109362), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IF
Human
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Rat
Expected
Tested
Expected

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

(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.

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

For unpurified use at 1/1000 - 1/10000.

Species

Rat

Dilution info

1/1000

Notes

For unpurified use at 1/1000 - 1/10000.

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/1000 - 1/10000.

Tested
Tested

Species

Human

Dilution info

1/150

Notes

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

2 products for Alternative Product

Target data

Function

Apoptosis-inducing protein that can overcome BCL2 suppression. May play a role in repartitioning calcium between the two major intracellular calcium stores in association with BCL2. Involved in mitochondrial quality control via its interaction with SPATA18/MIEAP: in response to mitochondrial damage, participates in mitochondrial protein catabolic process (also named MALM) leading to the degradation of damaged proteins inside mitochondria. The physical interaction of SPATA18/MIEAP, BNIP3 and BNIP3L/NIX at the mitochondrial outer membrane regulates the opening of a pore in the mitochondrial double membrane in order to mediate the translocation of lysosomal proteins from the cytoplasm to the mitochondrial matrix. Plays an important role in the calprotectin (S100A8/A9)-induced cell death pathway.

Alternative names

Recommended products

Anti-BNIP3 antibody [EPR4034] ab109362 is a rabbit monoclonal antibody that is used in BNIP3 western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with BNIP3 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR4034 is cited in over 80 publications

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR4034

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Stable for 12 months at -20°C

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

7 product images

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362), expandable thumbnail

    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, Anti-GAPDH antibody [mAbcam 9484] - 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 Anti-GAPDH antibody [mAbcam 9484] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

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

    Predicted band size: 21 kDa

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362), expandable thumbnail

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

    Blocking and diluting buffer: 5% NFDM/TBST.

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

    Lane 1: Rat kidney tissue lysate at 20 µg

    Lane 2: Mouse kidney tissue lysate at 20 µg

    Lane 3: Mouse spleen tissue lysate at 20 µg

    Secondary

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

    Predicted band size: 21 kDa

    Observed band size: 30 kDa

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362), expandable thumbnail

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

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

    Lane 1: Jurkat cell lysate treated with etoposide at 10 µg

    Lane 2: Jurkat cell lysate at 10 µg

    Lane 3: SH-SY5Y cell lysate treated with camptothecin at 10 µg

    Lane 4: SH-SY5Y cell lysate at 10 µg

    Lane 5: MCF-7 cell lysate treated with Cocl2 at 10 µg

    Predicted band size: 21 kDa

    Observed band size: 30 kDa

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

    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. Goat Anti-Rabbit IgG H&L (HRP) 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), expandable thumbnail

    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), expandable thumbnail

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

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

  • Western blot - Anti-BNIP3 antibody [EPR4034] (ab109362), expandable thumbnail

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

    The observed band sizes are consistent with publications. Bands around 30 and 60kDa should be monomer and dimer, respectively. Other bands should be different migrating forms (PMID: 15375294, PMID: 36396625, PMID: 9396766, PMID: 36396625, PMID: 33459136 etc.).
    1% SDS hot lysis method could help to get stronger monomer forms.

    The protocol used for this test is available under the Support & downloads section

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

    Lane 1: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 20ug, prepared by RIPA method at 20 µg

    Lane 2: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 20ug, prepared by 1% SDS hot method at 20 µg

    Secondary

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

    Exposure time: 156s

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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