Anti-Nrf2 抗体 (ab137550)
Key features and details
- Rabbit polyclonal to Nrf2
- Suitable for: IP, ChIP, WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-Nrf2 antibody
Nrf2 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Nrf2 -
由来種
Rabbit -
アプリケーション
適用あり: IP, ChIP, WB, ICC/IF, IHC-Pmore details -
種交差性
交差種: Human
交差が予測される動物種: Cow -
免疫原
Recombinant fragment corresponding to Human Nrf2 aa 108-413 (internal sequence).
Database link: Q16236 -
ポジティブ・コントロール
- WB: MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+). NRF2-transfected (+, including 3xFlag-tag) and HEK-293T whole cell extracts. ICC/IF: HeLa cells. IHC: Human breast carcinoma tissue. ChIP: HepG2 chromatin extract. IP: HepG2 whole cell extracts.
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特記事項
NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm.
Please note:
Nrf2 antibody (ab137550) detects no signal in most untreated samples for WB. Stimuli treated samples are recommended. Nrf2 expression is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109).
WB protocol info included in legend for MDA-MB-231 nuclear extract treated with 30 μM tBHQ for 4 hours (+).
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor®488 (ab150077). Or search our wide range of secondary antibodies for use with your experiment.The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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: 7.00
Preservative: 0.025% Proclin 300
Constituents: 78% PBS, 1% BSA, 20% Glycerol (glycerin, glycerine) -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Assay kits
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ChIP Related Products
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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ab137550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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IP |
1/100 - 1/500.
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ChIP |
Use at an assay dependent concentration.
Assay dependent |
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WB | (6) |
1/500 - 1/3000.
Detects a band of approximately 110 kDa (predicted molecular weight: 68 kDa). NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm. Stimuli treated samples are recommended. Expression levels may be better for nuclear fractions or for cell lines treated with MG132. |
ICC/IF |
1/100 - 1/1000.
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IHC-P | (1) |
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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特記事項 |
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IP
1/100 - 1/500. |
ChIP
Use at an assay dependent concentration. Assay dependent |
WB
1/500 - 1/3000. Detects a band of approximately 110 kDa (predicted molecular weight: 68 kDa). NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm. Stimuli treated samples are recommended. Expression levels may be better for nuclear fractions or for cell lines treated with MG132. |
ICC/IF
1/100 - 1/1000. |
IHC-P
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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: 497024 Cow
- Entrez Gene: 4780 Human
- Omim: 600492 Human
- SwissProt: Q5NUA6 Cow
- 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 (ab137550) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : NFE2L2 knockout A549 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : Huh7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 85 kDa why is the actual band size different from the predicted?Western blot: Anti-NFE2L2 antibody (ab137550) 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, ab137550 was shown to bind specifically to NFE2L2. A band was observed at 85 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. 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 (ab137550) at 1/500 dilution
Lane 1 : Untreated HepG2 whole cell extracts
Lane 2 : Treated HepG2 whole cell extracts
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : A HRP-conjugated anti-rabbit IgG antibody
Observed band size: 110 kDa why is the actual band size different from the predicted?The observed MW (110kDa) is based on PMID: 22703241
Various whole cell extracts (30 μg) were separated by 7.5% SDS-PAGE
Detection: Trident ECL plus-Enhanced.
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Paraffin embedded human breast carcinoma tissue stained for NRF2 using ab137550 at 1/500 dilution in immunohistochemical analysis.
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ab137550 staining Nrf2 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde. Samples were incubated with primary antibody diluted at 1/500 (green). Phalloidin, a cytoskeleton marker, diluted at 1/100 (red) was used.
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ChIP was performed with HepG2 chromatin extract and 5 μg of either normal rabbit IgG or Anti-Nrf2 antibody (ab137550). The precipitated DNA was detected by PCR with primer set targeting to GCLC gene locus.
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All lanes : Anti-Nrf2 antibody (ab137550) at 1/1000 dilution
Lane 1 : Untreated (-) MDA-MB-231 nuclear extract
Lane 2 : MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+)
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution
Developed using the ECL technique.
Observed band size: 110 kDa why is the actual band size different from the predicted?7.5% gel.
Running condition: 80V, 15min; 140V, 40min.
Transfer condition:Semi-dry, 18 V, 60min (Nitrocellulose membrane)
Blocking condition: 5% non-fat milk in TBST, RT, 60min.
Primary antibody incubation: 4°C overnight.
Secondary antibody incubation: Room temperature for 1 hour.
Washing condition: 5 ml TBST, 4 x 5min.
ECL detection.
The observed MW (110kDa) is based on PMID: 22703241
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ab137550 at 5 μg immunoprecipitating Nrf2 in HepG2 whole cell extracts.
Lane 1 (-): Ctrl IgG instead of ab137550 in HepG2 whole cell extracts.
Lane 2 (+): ab137550 + HepG2 whole cell extracts.For western blotting an anti-Rabbit IgG was used as a secondary reagent.
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All lanes : Anti-Nrf2 antibody (ab137550) at 1/1000 dilution
Lane 1 : Non-transfected (-) HEK-293T whole cell extracts
Lane 2 : NRF2-transfected (+, including 3xFlag-tag) HEK-293T whole cell extracts
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody
Observed band size: 110 kDa why is the actual band size different from the predicted?5% SDS-PAGE
The observed MW (110kDa) is based on PMID: 22703241
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 (335)
ab137550 は 335 報の論文で使用されています。
- Tang R et al. Sulforaphane activates anti-inflammatory microglia, modulating stress resilience associated with BDNF transcription. Acta Pharmacol Sin 43:829-839 (2022). PubMed: 34272506
- Chen T et al. Multiple myeloma cells depend on the DDI2/NRF1-mediated proteasome stress response for survival. Blood Adv 6:429-440 (2022). PubMed: 34649278
- Fu R et al. Neuroprotective Effects of Tetrahydroxystilbene Glucoside against Rotenone-Induced Toxicity in PC12 Cells. Biol Pharm Bull 45:143-149 (2022). PubMed: 34707025
- Gwak SY et al. Potential Role of Heme Oxygenase-1 in the Resolution of Experimentally Induced Colitis through Regulation of Macrophage Polarization. Gut Liver 16:246-258 (2022). PubMed: 34737242
- Beà A et al. Cardiac fibroblasts display endurance to ischemia, high ROS control and elevated respiration regulated by the JAK2/STAT pathway. FEBS J 289:2540-2561 (2022). PubMed: 34796659