Antibodies, Proteins, Kits and Reagents for Life Science

341 740 ₸ Product size

100 µl 341 740 ₸

Order now and get it on Tuesday March 02, 2021

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

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.

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

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)

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