Anti-Nrf2 (phospho S40) 抗体 [EP1809Y] (ab76026)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1809Y] to Nrf2 (phospho S40)
- Suitable for: Dot blot, ICC/IF, IHC-P, Flow Cyt (Intra), WB
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Nrf2 (phospho S40) antibody [EP1809Y]
Nrf2 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1809Y] to Nrf2 (phospho S40) -
由来種
Rabbit -
アプリケーション
適用あり: Dot blot, ICC/IF, IHC-P, Flow Cyt (Intra), WBmore details
適用なし: 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 asab133404) -
ポジティブ・コントロール
- WB: HepG2 whole cell lysate (ab7900). IHC-P: Human tonsil, breast carcinoma, ovarian carcinoma and cervical carcinoma tissue. ICC/IF: HepG2 cells. Flow Cyt (intra): Jurkat cells.
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特記事項
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
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
EP1809Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
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ab76026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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Dot blot |
Use at an assay dependent concentration.
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ICC/IF | (1) |
1/100 - 1/250.
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IHC-P | (3) |
1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/80 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (13) |
1/5000 - 1/50000. Predicted molecular weight: 68 kDa.
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特記事項 |
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Dot blot
Use at an assay dependent concentration. |
ICC/IF
1/100 - 1/250. |
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/80 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/5000 - 1/50000. 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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Nrf2 was abundantly expressed in carcinomas, low grade dysplasias, and non-atypical epithelia of oral tissue.
Representative findings of Nrf2 staining in carcinoma (left), in low grade dysplasia (middle), and in non-atypical epithelium (right).
Corresponding PLA signals are displayed in the lower row. Scale bar; 100 µm.
Surgical specimens were transferred to 10% buffered formalin and fixed overnight. The fixed samples were embedded in paraffin, and serially sliced into 5 µm sections. After dewaxing, sections were autoclaved at 120°C for 1 min in 10 mM sodium citrate buffer (pH 6.0), and immersed in 0.3% H2O2. They were then incubated overnight at 4°C with primary antibody to Nrf2 (diluted 1:200). The sections were rinsed with 1×PBS and incubated with the secondary antibody conjugated with horseradish peroxidase at room temperature for 1 hour. The sections were then stained with 3.3′-diaminobenzidinetetrahydrochloride (DAB) and counterstained with hematoxylin.
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Immunofluorescence analysis of Nrf2 levels in Kaposi's sarcoma skin lesions.
B) Healthy skin (top two rows) and KS skin tissue (bottom row) were double-stained for LANA-1 (Alexa-Fluor 594- red) and host phosphorylated pNrf2 (ab76026) (Alexa-Fluor®488 – green). DAPI was used to visualize the nuclei, and the triple merge of LANA-1, pNrf2 and DAPI is shown in the third column.
Yellow square = enlarged area.
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Nrf2 Translocation from cytoplasm to nucleus.
(A) Human islets were treated with dh404 for 0.5, 1 or 2 hours. The treated and untreated samples were stained with Nrf2 antibody ab76026 (Green) and DAPI (Blue). The con-focal microscope clearly showed that the Nrf2 translocation from cytoplasm to nucleus in the dh404 treated human islet cells
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All lanes : Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/50000 dilution (purified)
Lane 1 : untreated HepG2 cell lysate
Lane 2 : HepG2 treated with phosphatase lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 68 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab76026 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunofluorescence staining of HepG2 cells with purified ab76026 at a working dilution of 1/100, counter-stained with DAPI. The treated cells were treated with alkaline phosphatase for 1 h at 37°C. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab76026 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab76026 at a dilution of 1 in 80 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
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Dot blot analysis of Nrf2 peptides using unpurified ab76026 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated secondary antibody at 1/1000 dilution. Blocking and diluting buffer was 5% NFDM/TBST.
Lane 1: Nrf2 (pS40) phospho peptide
Lane 2: Nrf2 non-phospho peptide
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma using unpurified ab76026 at 1/100 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/10000 dilution (unpurified)
Lane 1 : Untreated HepG2 (human hepatocellular carcinoma) whole cell lysates 20µg
Lane 2 : HepG2 (human hepatocellular carcinoma) treated with Alkaline Phosphatase (AP) whole cell lysates 20µg.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 68 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST, dilution buffer: 5% NFDM /TBST, exposure time: 15 seconds
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All lanes : Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/20000 dilution (unpurified)
Lane 1 : HepG2 cell lysate
Lane 2 : HepG2 cell lysate treated with AP
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/1000 dilution
Predicted band size: 68 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted? -
Unpurified ab76026 staining Nrf2 (phospho S40) in Human normal lung tissue sections by IHC-P (Formaldehyde-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% casein for 30 minutes at 4°C. Antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/50) in 1% casein for 24 hours at 4°C. An undiluted HRP-conjugated Goat polyclonal to rabbit IgG was used as the secondary antibody.
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Unpurified ab76026 showing positive staining in Breast carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Unpurified ab76026 showing positive staining in Cervical carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Unpurified ab76026 showing positive staining in Ovarian carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Unpurified ab76026 showing positive staining in Normal tonsil tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (180)
ab76026 は 180 報の論文で使用されています。
- Ding P et al. BMPER alleviates ischemic brain injury by protecting neurons and inhibiting neuroinflammation via Smad3-Akt-Nrf2 pathway. CNS Neurosci Ther 28:593-607 (2022). PubMed: 34904361
- Wang CY et al. Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ2 Attenuates H2O2-Induced IL-6 Expression in Rat Brain Astrocytes. Neurotox Res 40:154-172 (2022). PubMed: 34997457
- Santini SJ et al. Copper-catalyzed dicarbonyl stress in NAFLD mice: protective effects of Oleuropein treatment on liver damage. Nutr Metab (Lond) 19:9 (2022). PubMed: 35148806
- Lee JJ et al. Galangin Reverses H2O2-Induced Dermal Fibroblast Senescence via SIRT1-PGC-1α/Nrf2 Signaling. Int J Mol Sci 23:N/A (2022). PubMed: 35163314
- Li M et al. Four New Benzoylamide Derivatives Isolated from the Seeds of Lepidium apetalum Willd. and Ameliorated LPS-Induced NRK52e Cells via Nrf2/Keap1 Pathway. Molecules 27:N/A (2022). PubMed: 35163986