Anti-Nrf2 antibody [EP1808Y] - Carrier free (ab180845)

Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845) is a rabbit recombinant monoclonal antibody provided in a PBS only buffer for easy conjugation detecting Nrf2 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, ICC/IF. Suitable for Human.

- Clone EP1808Y is the most cited clone to Nrf2

- KO validated for confirmed specificity

- BSA, sodium azide, and glycerol-free for easy conjugation

- Biophysical QC for unrivalled batch-batch consistency

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Images

Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (AB180845), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	ChIP	WB	Flow Cyt (Intra)	IHC-P
Human	Tested	Not recommended	Not recommended	Tested	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information NFE2L2

Function

The protein expressed by the NFE2L2 gene functions as a transcription factor that plays a key role in responding to oxidative stress by binding to antioxidant response elements (ARE) in the promoter regions of various cytoprotective genes, such as phase 2 detoxifying enzymes, to promote their expression and neutralize reactive electrophiles. Under normal conditions, it is ubiquitinated and degraded in the cytoplasm by the BCR(KEAP1) complex. However, oxidative stress leads to the inhibition of the BCR(KEAP1) complex, allowing NFE2L2/NRF2 to accumulate in the nucleus, form heterodimers with small Maf proteins, and bind to ARE elements. Additionally, the NFE2L2/NRF2 pathway is activated by selective autophagy, where KEAP1 interacts with SQSTM1/p62, inactivating the BCR(KEAP1) complex and leading to nuclear NFE2L2/NRF2 accumulation and cytoprotective gene expression. It may also be involved in activating genes of the beta-globin cluster by facilitating enhancer activity of hypersensitive site 2 of the beta-globin locus control region. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

NRF2, NFE2L2, Nuclear factor erythroid 2-related factor 2, NF-E2-related factor 2, NFE2-related factor 2, Nrf-2

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EP1808Y
Purification technique: Affinity purification Protein A
Specificity: The expression of Nrf2 is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109). Nrf2 antibody (ab62352) detects no signal in most untreated samples in WB. Stimuli treated samples are recommended. We do not recommend using this product in western blot with tissue lysates, however some customers have used this antibody successfully using concentrated samples (see submitted abreviews).
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

What is this antibody validated in?

Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunocytochemistry/immunofluorescence (ICC/IF) in Human samples.

What is the molecular weight of Nrf2?

Anti-Nrf2 [EP1808Y] - BSA and Azide free (ab180845) specifically detects a band for Nrf2 (UniProt: Q16236) at a molecular weight of 68kDa.

Trusted by the scientific community

Anti-Nrf2 [EP1808Y] - BSA and Azide free (ab180845) was first used in a scientific publication in 2016 and has been cited over 10 times in peer-reviewed journals.

Specificity confirmed

The specificity of Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845) has been confirmed by Western blot testing in NFE2L2 Knockout A549 cell line, ab285359.

Other related products

We have a range of other formats of antibody clone [EP1808Y] also available for your convenience:
ab62352, Carrier free - ab180845, Alexa Fluor® 488 - ab194984, Alexa Fluor® 647 - ab194985, Alexa Fluor® 594 - ab206890, PE - ab223926, APC - ab223927

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Nrf2 also known as nuclear factor erythroid 2-related factor 2 is a transcription factor with a molecular weight of approximately 66 kDa. It plays a mechanical role in regulating the expression of antioxidant proteins that protect against oxidative damage triggered by injury and inflammation. Nrf2 is ubiquitously expressed in various tissues including the liver lungs and skin. Nrf2 activation occurs when it translocates from the cytoplasm to the nucleus to bind the antioxidant response element (ARE) in the DNA starting transcription of target genes.

Biological function summary

Nrf2 acts as an important regulator of the cellular antioxidant response. It works in conjunction with its partner protein Keap1 forming a complex that controls its stability and degradation. Under normal conditions Keap1 keeps Nrf2 in the cytoplasm where it is targeted for ubiquitination and degradation. Once activated by oxidative stress or electrophiles Nrf2 dissociates from Keap1 thereby avoiding degradation and relocates to the nucleus to activate the transcription of ARE-dependent genes. This activity boosts the cellular response to oxidative stress by inducing genes involved in detoxification and cellular defense.

Pathways

Nrf2 plays a significant role in the oxidative stress response and detoxification pathways. Nrf2 activation is linked closely to the PI3K/Akt signaling pathway which influences cell survival growth and metabolism. This pathway also interacts with other important proteins like GSK-3Β which can modulate Nrf2 activity and stability. Through these pathways Nrf2 orchestrates a defense mechanism against reactive oxygen species (ROS) by boosting the expression of antioxidant enzymes and detoxifying proteins.

Associated diseases and disorders

Nrf2 has been associated with conditions like cancer and neurodegenerative diseases. In cancer aberrant Nrf2 activation may lead to enhanced tumor survival by increasing expression of cytoprotective genes which makes cancer cells resistant to chemotherapy. Nrf2 also interacts with proteins such as p53 which play roles in tumor suppression and cellular stress responses. In neurodegenerative disorders reduced Nrf2 activity can contribute to oxidative stress leading to neuron damage and disease progression with proteins like amyloid-beta also being linked with oxidative processes affected by Nrf2 functionality.

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:
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10 product images

Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-Nrf2 antibody [EP1808Y] ab194985 for protocol details.
Alexa Fluor® 647 Anti-Nrf2 antibody [EP1808Y] ab194985 staining Nrf2 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 647 Anti-Nrf2 antibody [EP1808Y] ab194985 at a working dilution 1/100 (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
This product gave a positive signal in 4% formaldehyde (10 min) fixed HeLa cells under the same testing conditions.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Flow Cytometry (Intracellular) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (PE). Please refer to PE Anti-Nrf2 antibody [EP1808Y] ab223926 for protocol details.
Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with PE Anti-Nrf2 antibody [EP1808Y] ab223926 (red line). The cells were fixed with 4% formaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Triton X-100 for 15 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (PE Anti-Nrf2 antibody [EP1808Y] ab223926, 1/5000 dilution) for 30 minutes at 22°C.
Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.
This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Triton X-100 for 15 minutes used under the same conditions.
Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-Nrf2 antibody [EP1808Y] ab194984 for protocol details.
Alexa Fluor® 488 Anti-Nrf2 antibody [EP1808Y] ab194984 staining Nrf2 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 488 Anti-Nrf2 antibody [EP1808Y] ab194984 at a working dilution of 1/100 (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 594, shown in red) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Nrf2 with purified Anti-Nrf2 antibody [EP1808Y] ab62352 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. The cells were co-stained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, an Alexa Fluor^® 594-conjugated mouse anti-alpha tubulin antibody (1/200). Nuclei counterstained with DAPI (blue).
Secondary antibody only control: PBS was used instead of the primary antibody as the negative control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Nrf2 antibody [EP1808Y] ab62352).
Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Nrf2 with purified Anti-Nrf2 antibody [EP1808Y] ab62352 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody (1/1000) was used as the secondary antibody. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594). DAPI was used to stain the nuclei blue.
Secondary antibody only control: PBS was used instead of the primary antibody as the negative control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Nrf2 antibody [EP1808Y] ab62352).
Flow Cytometry (Intracellular) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Intracellular Flow Cytometry analysis of HeLa cells labelling Nrf2 with purified Anti-Nrf2 antibody [EP1808Y] ab62352 at a dilution of 1/60 (red). Cells were fixed with 4% paraformaldehyde. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Nrf2 antibody [EP1808Y] ab62352).
Flow Cytometry (Intracellular) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Nrf2 with Anti-Nrf2 antibody [EP1808Y] ab62352 at 1/40 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor^®488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Nrf2 antibody [EP1808Y] ab62352).
Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
This data was developed using Anti-Nrf2 antibody [EP1808Y] ab62352, the same antibody clone in a different buffer formulation.
Anti-Nrf2 antibody [EP1808Y] ab62352 staining Nrf2 in untreated wild type A549 cells (left panel), treated wild type A549 cells (middle panel) and untreated NFE2L2 knockout A549 cells (right panel). Cells were treated with 2µM of MG-132 for 18 hours (MG-132, proteasome inhibitor ab141003). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Nrf2 antibody [EP1808Y] ab62352 at 0.2 µg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2µg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2µg/ml (shown in magenta). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Western blot - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
All lanes: Western blot - Anti-Nrf2 antibody [EP1808Y] (Anti-Nrf2 antibody [EP1808Y] ab62352) at 1/500 dilution
Lane 1: Western blot - Human NFE2L2 (Nrf2) knockout HeLa cell lysate (Human NFE2L2 (Nrf2) knockout HeLa cell lysate ab263934)
Lane 1: Wild-type HeLa control MG132 (0 uM, 18 h) cell lysate
Lane 2: Wild-type HeLa treated MG132 (2 uM, 18 h) cell lysate
Lane 2: Western blot - Human NFE2L2 (Nrf2) knockout HeLa cell line (Human NFE2L2 (Nrf2) knockout HeLa cell line ab262507)
Lane 3: NFE2L2 knockout HeLa control MG132 (0 uM, 18 h) cell lysate
Lane 4: NFE2L2 knockout HeLa treated MG132 (2 uM, 18 h) cell lysate
Lane 5: HCT 116 cell lysate
Western blot - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Nrf2 Western blot staining using rabbit Anti-Nrf2 antibody
This data was developed using Anti-Nrf2 antibody [EP1808Y] ab62352, the same antibody clone in a different buffer formulation.
False colour image of Western blot: Anti-Nrf2 antibody [EP1808Y] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Nrf2 antibody [EP1808Y] ab62352 was shown to bind specifically to Nrf2.
Target band (indicated by arrow) was observed at 85-90 kDa in wild-type A549 cell lysates with no signal observed at this size in NFE2L2 knockout cell line Human NFE2L2 knockout A549 cell line[21] ab285359 (knockout cell lysate ab289682). A band lower than the target was present on both WT and KO. According to the literature (PMID: 40043449), the lower band is calmegin.
To generate this image, wild-type and NFE2L2 knockout A549 cell lysates were analysed. Please note that MG132 treatment does not affect expression levels of Nrf2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Nrf2 antibody [EP1808Y] (Anti-Nrf2 antibody [EP1808Y] ab62352) at 1/1000 dilution
Lane 1: Wild-type A549 Vehicle control MG132 (0 uM, 18 h) cell lysate at 20 µg
Lane 2: Wild-type A549 Treated MG132 (2 uM, 18 h) cell lysate at 20 µg
Lane 3: NFE2L2 [21] knockout A549 Vehicle control MG132 (0 uM, 18 h) cell lysate at 20 µg
Lane 4: NFE2L2 [21] knockout A549 Treated MG132 (2 uM, 18 h) cell lysate at 20 µg
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 68 kDa