Anti-Nrf2 抗体 [EP1808Y] - BSA and Azide free (ab180845)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1808Y] to Nrf2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free
Nrf2 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1808Y] to Nrf2 - BSA and Azide free -
由来種
Rabbit -
特異性
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).
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アプリケーション
適用あり: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore details
適用なし: ChIP or IP -
種交差性
交差種: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab167152) -
ポジティブ・コントロール
- WB: MG-132 treated HeLa whole cell lysate, THP-1, MG-132 treated HepG2 whole cell lysate, MG-132 treated HCT-116 and A549 whole cell lysate. IHC-P: Human pancreatic carcinoma and kidney cancer tissues. ICC/IF: HepG2 and HeLa cells. Flow Cyt (intra): HeLa cells.
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特記事項
ab180845 is the carrier-free version of ab62352.
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.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
バッファー
Constituent: 100% PBS -
キャリア・フリー
はい -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1808Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Alexa Fluor® 488 Anti-Nrf2 antibody [EP1808Y] (ab194984)
- Alexa Fluor® 647 Anti-Nrf2 antibody [EP1808Y] (ab194985)
- HRP Anti-Nrf2 antibody [EP1808Y] (ab194986)
- Alexa Fluor® 594 Anti-Nrf2 antibody [EP1808Y] (ab206890)
- PE Anti-Nrf2 antibody [EP1808Y] (ab223926)
- APC Anti-Nrf2 antibody [EP1808Y] (ab223927)
- Anti-Nrf2 antibody [EP1808Y] (ab62352)
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Conjugation kits
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Isotype control
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab180845の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa.
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特記事項 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa. |
ターゲット情報
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機能
Transcription activator that binds to antioxidant response (ARE) elements in the promoter regions of target genes. Important for the coordinated up-regulation of genes in response to oxidative stress. May 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. -
組織特異性
Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle. -
配列類似性
Belongs to the bZIP family. CNC subfamily.
Contains 1 bZIP domain. -
ドメイン
Acidic activation domain in the N-terminus, and DNA binding domain in the C-terminus. -
翻訳後修飾
Phosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus. -
細胞内局在
Cytoplasm > cytosol. Nucleus. Cytosolic under unstressed conditions, translocates into the nucleus upon induction by electrophilic agents. - Information by UniProt
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参照データベース
- Entrez Gene: 4780 Human
- Omim: 600492 Human
- SwissProt: Q16236 Human
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別名
- erythroid derived 2 antibody
- HEBP1 antibody
- like 2 antibody
see all
画像
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All lanes : 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
Lysates/proteins at 20 µg per lane.
Predicted band size: 68 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Nrf2 antibody [EP1808Y] - ChIP Grade staining at 1/500 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. A band was observed at 85 kDa in wild-type HeLa cell lysates with no signal observed at this size in NFE2L2 CRISPR-Cas9 edited cell line ab262507 (CRISPR-Cas9 edited cell lysate ab263934). The band observed in the CRISPR-Cas9 edited lysate lane below 85 kDa is likely to represent a truncated form of Nrf2. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and NFE2L2 CRISPR-Cas9 edited HeLa cell lysates were analysed. 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/1000 dilution
Lane 1 : Wild-type A549 Vehicle control MG132 (0 uM, 18 h) cell lysate
Lane 2 : Wild-type A549 Treated MG132 (2 uM, 18 h) cell lysate
Lane 3 : NFE2L2 [21] knockout A549 Vehicle control MG132 (0 uM, 18 h) cell lysate
Lane 4 : NFE2L2 [21] knockout A549 Treated MG132 (2 uM, 18 h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 85-90 kDa why is the actual band size different from the predicted?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. A band 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). 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.
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All lanes : Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate treated with MG-132 2uM for 18h
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 68 kDa
Observed band size: 955-110 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThis data was developed using ab62352, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concnetration: 5% NFDM/TBST.
ab181602 was used as GAPDH loading control.
The two bands are different isoforms of Nrf2, the molecular weight observed is consistent with what has been described in the literature: PMID: 17512459, PMID: 22703241.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreatic carcinoma tissue labelling Nrf2 with purified ab62352 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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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).
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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.
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Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 488). Please refer to ab194984 for protocol details.
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 ab194984 at a working dilution of 1/100 (shown in green) and 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).
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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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney cancer tissue sections labeling Nrf2 with ab62352 at 1/100 dilution. The tissue was fixed with paraformaldehyde and a heat mediated antigen retrival step was performed with TRIS-EDTA Buffer pH 9.0. Staining with ab62352 at 1/100 was carried out in a dilution buffer with blocking for 30 minutes at 20°C. A undiluted goat anti-rabbit HRP conjugated secondary antibody was used.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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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).
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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).
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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).
プロトコール
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (13)
ab180845 は 13 報の論文で使用されています。
- Chhunchha B et al. Switching of Redox Signaling by Prdx6 Expression Decides Cellular Fate by Hormetic Phenomena Involving Nrf2 and Reactive Oxygen Species. Cells 11:N/A (2022). PubMed: 35455944
- Bu X et al. CD147 confers temozolomide resistance of glioma cells via the regulation of β-TrCP/Nrf2 pathway. Int J Biol Sci 17:3013-3023 (2021). PubMed: 34421346
- Chhunchha B et al. Sulforaphane-Induced Klf9/Prdx6 Axis Acts as a Molecular Switch to Control Redox Signaling and Determines Fate of Cells. Cells 8:N/A (2019). PubMed: 31569690
- Hu T et al. Clinicopathologic significance of CXCR4 and Nrf2 in colorectal cancer. J Biomed Res 27:283-90 (2013). IHC ; Human . PubMed: 23885267
- Ma J et al. PALB2 interacts with KEAP1 to promote NRF2 nuclear accumulation and function. Mol Cell Biol 32:1506-17 (2012). WB . PubMed: 22331464