リコンビナント
RabMAb

Anti-Nrf2 (phospho S40) 抗体 [EP1809Y] (ab76026)

製品の概要

  • 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)
  • アプリケーション
    適用あり: ICC/IF, IHC-P, WB, Flow Cytmore details
    適用なし: IP
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic phospho-peptide (Human) corresponding to residues near serine 40 of Nrf2.

  • ポジティブ・コントロール
    • Human breast carcinoma tissue, HepG2 cell lysate
  • 特記事項

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

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    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 ab76026 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
ICC/IF 1/100 - 1/250.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

WB 1/5000 - 1/50000. Predicted molecular weight: 68 kDa.
Flow Cyt 1/80 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • 追加情報
    Is unsuitable for IP.
  • 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

    Anti-Nrf2 (phospho S40) antibody [EP1809Y] 画像

    • 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
      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

    • 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).
    • 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.

    • 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.
    • 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

    • 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
      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

    • 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
      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?)
    • Immunohistochemical analysis of paraffin-embedded human breast carcinoma using unpurified ab76026 at 1/100 dilution.

    • 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.

      See Abreview

    • Unpurified ab76026 showing positive staining in Breast carcinoma tissue.

    • Unpurified ab76026 showing positive staining in Cervical carcinoma tissue.

    • Unpurified ab76026 showing positive staining in Ovarian carcinoma tissue.

    • Unpurified ab76026 showing positive staining in Normal tonsil tissue.

    Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) 使用論文

    This product has been referenced in:
    • Arora R  et al. Esculetin induces antiproliferative and apoptotic response in pancreatic cancer cells by directly binding to KEAP1. Mol Cancer 15:64 (2016). WB ; Human . Read more (PubMed: 27756327) »
    • Wu ST  et al. Cellular effects induced by 17-ß-estradiol to reduce the survival of renal cell carcinoma cells. J Biomed Sci 23:67 (2016). WB ; Human . Read more (PubMed: 27680214) »

    See all 27 Publications for this product

    Product Wall

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (pancreas)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10 mM TRIS-EDTA pH 9 at 98°C for 20 minutes, normal pressure
    Permeabilization
    Yes - 0.04% triton 100 x in the blocking solution
    Specification
    pancreas
    Blocking step
    BSA as blocking agent for 15 minute(s) · Concentration: 2% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Dec 13 2016

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (lungs)
    Gel Running Conditions
    Reduced Denaturing (30 minutes RT, 10% gel)
    Loading amount
    100 µg
    Specification
    lungs
    Blocking step
    2 % Topblock in TBT-0.1% Triton 100 x as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    投稿 Dec 13 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (brain)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    50 µg
    Specification
    brain
    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 Jul 12 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Specification
    Brain
    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 Jul 12 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (breast cancer cell line xenograft)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
    Permeabilization
    No
    Specification
    breast cancer cell line xenograft
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Dec 17 2015

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human Placenta)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    30 µg
    Specification
    Human Placenta
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Vicky

    Verified customer

    投稿 Nov 13 2015

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Skin)
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Fixative
    Acetone
    Username

    Abcam user community

    Verified customer

    投稿 Nov 09 2015

    ab76026 will detect Nrf2 phosphorylated at S40 regardless of whether or not it is in the nucleus. The understanding from the literature is that under some conditions Nrf2 is phosphorylated at S40 and this leads to translocation of Nrf2 from the cytopla...

    Read More
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (From muscle, liver, aorta)
    Gel Running Conditions
    Reduced Denaturing (4-20% gradient gel)
    Loading amount
    35 µg
    Specification
    From muscle, liver, aorta
    Blocking step
    Licor as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Username

    Dr. Thunder Jalili

    Verified customer

    投稿 May 14 2015

    Application
    Western blot
    Loading amount
    25 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Tissue lysate - whole (liver)
    Specification
    liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Abcam user community

    Verified customer

    投稿 Dec 19 2014

    1-10 of 26 Abreviews or Q&A

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

    登録