概述

  • 产品名称

    Anti-Nrf2抗体[EP1808Y] - ChIP Grade
    参阅全部 Nrf2 一抗
  • 描述

    兔单克隆抗体[EP1808Y] to Nrf2 - ChIP Grade
  • 宿主

    Rabbit
  • 特异性

    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.

  • 经测试应用

    适用于: IHC-Fr, Flow Cyt, ChIP, ICC/IF, WB, IP, IHC-Pmore details
  • 种属反应性

    与反应: 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)

  • 阳性对照

    • WB: MG-132 treated HeLa whole cell lysate, Saos-2, THP-1, MG-132 treated HepG2 whole cell lysate, MG-132 treated HCT-116 whole cell lysate. IHC-P: Human pancreatic carcinoma and kidney cancer tissues. ICC/IF: HepG2 and HeLa cells. Flow Cyt: HeLa cells. IP: SAOS-2 whole cell lysate. ChIP: Chromatin prepared from HepG2 cells.
  • 常规说明

    PLEASE NOTE:

    Nrf2 antibody (ab62352) 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)

    Mouse is not recommended for IHC (weak 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.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

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.

应用 Ab评论 说明
IHC-Fr 1/100. Fix with acetone. Note: antigen retrieval is recommended.
Flow Cyt 1/40.
ChIP Use 2 µg for 25 µg of chromatin.
ICC/IF 1/100 - 1/500.
WB 1/200 - 1/1000. Predicted molecular weight: 68 kDa.Can be blocked with Nrf2 peptide (ab167152).
IP 1/20.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

靶标

  • 功能

    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.
  • 组织特异性

    Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.
  • 序列相似性

    Belongs to the bZIP family. CNC subfamily.
    Contains 1 bZIP domain.
  • 结构域

    Acidic activation domain in the N-terminus, and DNA binding domain in the C-terminus.
  • 翻译后修饰

    Phosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus.
  • 细胞定位

    Cytoplasm > cytosol. Nucleus. Cytosolic under unstressed conditions, translocates into the nucleus upon induction by electrophilic agents.
  • Information by UniProt
  • 数据库链接

  • 别名

    • 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

图片

  • All lanes : Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (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.

  • 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 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.

  • All lanes : Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (ab62352) at 1/200 dilution (Purified)

    Lane 1 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 2 : HCT 116 (Human colorectal carcinoma epithelial cell) treated with 25uM MG-132 for 4 hours whole cell lysates with 5% NFDM/TBST

    Lysates/proteins at 20 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: 60 seconds


    The expression of Nrf2 is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109). ab62352 detects no signal in most of the untreated samples in WB. Stimuli treated samples are recommended.

  • 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.

  • All lanes : Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (ab62352) at 1/200 dilution (Purified)

    Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) treated with 2uM MG-132 for 18 hours whole cell lysates with 5% NFDM/TBST

    Lysates/proteins at 20 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: 180 seconds


    The expression of Nrf2 is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109). ab62352 detects no signal in most of the untreated samples in WB. Stimuli treated samples are recommended.

  • 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

  • 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.

  • 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 VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • 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.

  • 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.

文献

This product has been referenced in:

  • Wu B  et al. Nrf2 overexpression protects against paraquat-induced A549 cell injury primarily by upregulating P-glycoprotein and reducing intracellular paraquat accumulation. Exp Ther Med 17:1240-1247 (2019). Read more (PubMed: 30679998) »
  • Chen X  et al. Adropin protects against liver injury in nonalcoholic steatohepatitis via the Nrf2 mediated antioxidant capacity. Redox Biol 21:101068 (2019). Read more (PubMed: 30684890) »
See all 176 Publications for this product

客户评价及客户问答

1-10 of 40 Abreviews or Q&A

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

Mr. Rudolf Jung

Verified customer

提交于 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

Miss. Sarah Rodrigues Moreira

Verified customer

提交于 Mar 01 2013

Answer

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 testing data to spoort the use of this antibody in mouse however, so I am wondering if this antibody is reacting non-specifically in mouse samples. We do have ab31162 and ab92946 which are tested and guaranteed to work in mouse, these may be better antibodies for you to work with. Please let me know if I can be of further assistance.

Read More
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

Amelia Romoser

Verified customer

提交于 Jan 28 2011

Application
Western blot
Sample
Human Cell lysate - whole cell (human melanoma cells)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
40 µg
Treatment
7,5 µM Sulforaphane for 24hrs
Specification
human melanoma cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

提交于 Oct 04 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (C42B Prostate cancer cell line)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
C42B Prostate cancer cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. Dimitra Kalamida

Verified customer

提交于 May 14 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HUVECs)
Permeabilization
Yes - txri-100
Specification
HUVECs
Blocking step
quickblock liquid as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Mar 28 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (cancer cell lines)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
25 µg
Specification
cancer cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Dec 05 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEFs)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Treatment
2uM MG132 18hrs
Specification
MEFs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Aug 15 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (fibroblast)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (gel 10%)
Loading amount
30 µg
Specification
fibroblast
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Jun 25 2018

1-10 of 40 Abreviews or Q&A

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