Name: Anti-BNIP3 antibody [EPR4034]
Brand: RabMAb
SKU: ab109362
Rating: 4 (3 reviews)

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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

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Primary

Secondary

Protein

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Primary - Rabbit Anti-BNIP3 antibody [EPR4034] (ab109362)
WB, IHC-P, ICC/IF

Secondary - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
IHC-P, WB, ELISA, IP

Protein - Recombinant Mouse BNIP3 protein (His tag) (ab235015)
SDS-PAGE

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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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR4034
Isotype

IgG
Research areas

Alternative Versions
- Anti-BNIP3 antibody [EPR4034] - BSA and Azide free (ab219609)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor? 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Mouse BNIP3 protein (His tag) (ab235015)
Related Products

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

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.

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.
Target information above from: UniProt accession Q12983 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 664 Human
- Entrez Gene: 12176 Mouse
- Entrez Gene: 84480 Rat
- Omim: 603293 Human
- SwissProt: Q12983 Human
- SwissProt: O55003 Mouse
- Unigene: 144873 Human
- Unigene: 378890 Mouse
- Unigene: 2060 Rat
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

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)

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)

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)

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)

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) 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) 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 analysis of SH-SY5Y cells labelling BNIP3 with unpurified ab109362 at a 1/100 dilution.
Anti-BNIP3 antibody [EPR4034] (ab109362)

Western blot protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

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

ab109362 has been referenced in 28 publications.

Ma Z et al. BNIP3 decreases the LPS-induced inflammation and apoptosis of chondrocytes by promoting the development of autophagy. J Orthop Surg Res 15:284 (2020). PubMed: 32723351
Tang C et al. Activation of BNIP3-mediated mitophagy protects against renal ischemia-reperfusion injury. Cell Death Dis 10:677 (2019). PubMed: 31515472
Ren Q et al. 6-Gingerol protects cardiocytes H9c2 against hypoxia-induced injury by suppressing BNIP3 expression. Artif Cells Nanomed Biotechnol 47:2016-2023 (2019). PubMed: 31223035
Rémy M et al. Isolation and Culture of Human Stem Cells from Apical Papilla under Low Oxygen Concentration Highlight Original Properties. Cells 8:N/A (2019). PubMed: 31766521
Zhang J et al. Involvement of autophagy in hypoxia-BNIP3 signaling to promote epidermal keratinocyte migration. Cell Death Dis 10:234 (2019). PubMed: 30850584

View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Dec 27 2019

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

Antoine Chaanine

Verified customer

Submitted Mar 31 2017

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

Abcam user community

Verified customer

Submitted Feb 20 2017

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (28)

Customer reviews and Q&As

Western blot abreview for Anti-BNIP3 antibody [EPR4034]

Western blot abreview for Anti-BNIP3 antibody [EPR4034]

Western blot abreview for Anti-BNIP3 antibody [EPR4034]