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Knockout Tested Rabbit Recombinant Monoclonal Nrf2 antibody. Carrier free. Suitable for ICC/IF, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 13 publications.

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Images

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.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPChIPWBFlow Cyt (Intra)IHC-P
Human
Tested
Not recommended
Not recommended
Tested
Tested
Not recommended

Tested
Tested

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

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

Function

Transcription factor that plays a key role in the response to oxidative stress: binds to antioxidant response (ARE) elements present in the promoter region of many cytoprotective genes, such as phase 2 detoxifying enzymes, and promotes their expression, thereby neutralizing reactive electrophiles (PubMed:11035812, PubMed:19489739, PubMed:29018201, PubMed:31398338). In normal conditions, ubiquitinated and degraded in the cytoplasm by the BCR(KEAP1) complex (PubMed:11035812, PubMed:15601839, PubMed:29018201). In response to oxidative stress, electrophile metabolites inhibit activity of the BCR(KEAP1) complex, promoting nuclear accumulation of NFE2L2/NRF2, heterodimerization with one of the small Maf proteins and binding to ARE elements of cytoprotective target genes (PubMed:19489739, PubMed:29590092). The NFE2L2/NRF2 pathway is also activated in response to selective autophagy: autophagy promotes interaction between KEAP1 and SQSTM1/p62 and subsequent inactivation of the BCR(KEAP1) complex, leading to NFE2L2/NRF2 nuclear accumulation and expression of cytoprotective genes (PubMed:20452972). May also be involved in the transcriptional activation of genes of the beta-globin cluster by mediating enhancer activity of hypersensitive site 2 of the beta-globin locus control region (PubMed:7937919).

Alternative names

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Knockout Tested Rabbit Recombinant Monoclonal Nrf2 antibody. Carrier free. Suitable for ICC/IF, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 13 publications.

Alternative names

Key facts

Isotype

IgG

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
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Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab180845 is the carrier-free version of ab62352.

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

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.

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

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.

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.

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:
Terms & Conditions.

9 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845), expandable thumbnail

    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 ab194985 for protocol details.

    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 ab194985 at a working dilution 1/100 (shown in red) and 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), expandable thumbnail

    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 ab223926 for protocol details.

    Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with 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 (ab223926, 1/5000 dilution) for 30 minutes at 22°C.

    Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (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), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Nrf2 with purified ab62352 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. The cells were co-stained with 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 (ab62352).

  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Nrf2 with purified 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) secondary antibody (1/1000) was used as the secondary antibody. Cells were counterstained with 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 (ab62352).

  • Flow Cytometry (Intracellular) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Intracellular Flow Cytometry analysis of HeLa cells labelling Nrf2 with purified ab62352 at a dilution of 1/60 (red). Cells were fixed with 4% paraformaldehyde. 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 (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 (ab62352).

  • Flow Cytometry (Intracellular) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845), expandable thumbnail

    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 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 (ab62352).

  • Western blot - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845), expandable thumbnail

    Western blot - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    All lanes: Western blot - Anti-Nrf2 antibody [EP1808Y] (AB62352) at 1/500 dilution

    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 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

  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    This data was developed using ab62352, the same antibody clone in a different buffer formulation.

    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 (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 ab62352 at 0.2 µg/ml concentration and 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) (ab150081) at 2µg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (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), expandable thumbnail

    Western blot - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    This data was developed using 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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 ab285359 (knockout cell lysate ab289682). A band lower than the target was present on both WT and KO, we are unsure about the identity of this band.

    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] - ChIP Grade (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

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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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