リコンビナント
RabMAb

Anti-Nrf2 抗体 [EP1808Y] (ab62352)

製品の概要

  • 製品名
    Anti-Nrf2 antibody [EP1808Y]
    Nrf2 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP1808Y] to Nrf2
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: ChIP, IHC-Fr, Flow Cyt, ICC/IF, WB, IP, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide within Human Nrf2 aa 550 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q16236
    (Peptide available as ab167152)

  • ポジティブ・コントロール
    • WB: MG-132 treated HeLa whole cell lysate, Saos-2, THP-1 and HepG2 cell lysate. IHC-P: Human pancreatic carcinoma tissue. ICC/IF: HepG2 and HeLa cells. Flow Cyt: HeLa cells. ChIP: Chromatin from HepG2 cells. IP: SAOS-2 whole cell lysate.
  • 特記事項

    Mouse is not recommended for IHC for weakly staining.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab62352 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ChIP Use at an assay dependent concentration. PubMed: 22581777
IHC-Fr 1/100. Fix with acetone. Note: antigen retrieval is recommended.
Flow Cyt 1/40.
ICC/IF 1/100 - 1/500.
WB 1/1000 - 1/5000. Predicted molecular weight: 68 kDa.Can be blocked with Nrf2 peptide (ab167152).
IP 1/20.
IHC-P 1/100 - 1/250.

ターゲット情報

  • 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
  • 参照データベース
  • 別名
    • erythroid derived 2 antibody
    • HEBP1 antibody
    • like 2 antibody
    • NF E2 related factor 2 antibody
    • NF-E2-related factor 2 antibody
    • NF2L2_HUMAN antibody
    • NFE2 related factor 2 antibody
    • NFE2-related factor 2 antibody
    • Nfe2l2 antibody
    • Nrf 2 antibody
    • NRF2 antibody
    • Nuclear factor (erythroid derived 2) like 2 antibody
    • Nuclear factor antibody
    • nuclear factor erythroid 2 like 2 antibody
    • Nuclear factor erythroid 2 related factor 2 antibody
    • Nuclear factor erythroid 2-related factor 2 antibody
    • Nuclear factor erythroid derived 2 like 2 antibody
    see all

画像

  • All lanes : Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/1000 dilution (purified)

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HeLa whole cell lysate treated with MG-132 2 µM for 18 hours
    Lane 3 : Saos-2 whole cell lysate
    Lane 4 : THP-1 whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 68 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure:
    Lanes 1 and 2: 15 seconds.
    Lanes 3 and 4: 3 minutes.

  • All lanes : Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/1000 dilution (purified)

    Lane 1 : MEF (Mouse embryonic fibroblast) whole cell lysates
    Lane 2 : MEF (Mouse embryonic fibroblast) treated with MG-132 2uM for 18 hours whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 68 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST

  • 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.

  • Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/2000 dilution (unpurified) + HepG2 cell lysate at 10 µg

    Secondary
    goat anti rabbit HRP at 1/2000 dilution

    Predicted band size: 68 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)

  • ab62352 (purified) at 1/30 immunoprecipitating Nrf2 in SAOS-2 whole cell lysate.

    Lane 1 (input): SAOS-2 whole cell lysate (10µg)
    Lane 2 (+): ab62352 + SAOS-2 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab62352 in SAOS-2 whole cell lysate.

    For western blotting, ab62352 was used at a dilution of 1/500 and ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1000).

    Blocking and dilution buffer: 5% NFDM/TBST.

  • 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.

  • 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.

  • Chromatin was prepared from HepG2 cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab62352 (blue), and 20µl of A/G sepharose bead slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • 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.

  • 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.  

    See Abreview

  • 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.

参考文献

This product has been referenced in:
  • Antognelli C  et al. KRIT1 loss-of-function induces a chronic Nrf2-mediated adaptive homeostasis that sensitizes cells to oxidative stress: Implication for Cerebral Cavernous Malformation disease. Free Radic Biol Med 115:202-218 (2018). Read more (PubMed: 29170092) »
  • Antognelli C  et al. Data in support of sustained upregulation of adaptive redox homeostasis mechanisms caused by KRIT1 loss-of-function. Data Brief 16:929-938 (2018). Read more (PubMed: 29511711) »

See all 84 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
Sample
Human Tissue sections (kidney cancer)
Specification
kidney cancer
Permeabilization
Yes - Wash Buffer with Tween
Fixative
Paraformaldehyde
Username

Mr. Rudolf Jung

Verified customer

投稿 Aug 23 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Bone Marrow derived macrophages (BMDM))
Specification
Bone Marrow derived macrophages (BMDM)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.2% Triton
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Miss. Sarah Rodrigues Moreira

Verified customer

投稿 Mar 01 2013

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results. We have not had a band reported to use of this size, the predicted MW is around 68 kD but we saw the band a bit higher at 100 kD. We do not have t...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (A549)
Loading amount
15 µg
Specification
A549
Treatment
50 uM BSO
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Amelia Romoser

Verified customer

投稿 Jan 28 2011

Application
Western blot
Sample
Mouse Tissue lysate - whole (B16 tumor lysate)
Gel Running Conditions
Reduced Denaturing (4-20% TG gels)
Loading amount
50 µg
Specification
B16 tumor lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Apr 10 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MDA-MB-231 cells)
Gel Running Conditions
Reduced Denaturing (4-20% TG gels)
Loading amount
50 µg
Specification
MDA-MB-231 cells
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Apr 10 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (lung)
Gel Running Conditions
Reduced Denaturing (4-15% gradient gel)
Loading amount
20 µg
Specification
lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Username

Abcam user community

Verified customer

投稿 Sep 19 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (ARPE-19)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Treatment
cigarette smoke for 0.5-24 hrs
Specification
ARPE-19
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. lei wang

Verified customer

投稿 Jul 07 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Human Mammary Epithelial Cells (HMEC))
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
10000 cells
Specification
Human Mammary Epithelial Cells (HMEC)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Aug 22 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain tissue)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
50 µg
Specification
Brain tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 May 31 2016

1-10 of 34 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録