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RabMAb

Anti-NADPH oxidase 4 抗体 [UOTR1B493] (ab133303)

製品の概要

  • 製品名
    Anti-NADPH oxidase 4 antibody [UOTR1B493]
    NADPH oxidase 4 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [UOTR1B493] to NADPH oxidase 4
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: IHC-Fr, Flow Cyt, WB, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Rat, Human
  • 免疫原

    Synthetic peptide within Human NADPH oxidase 4. The exact sequence is proprietary.
    Database link: Q9NPH5
    (Peptide available as ab155071)

  • ポジティブ・コントロール
    • WB: Human fetal kidney tissue lysate, U87-MG, 293 and JAR cell lysates. IHC-P: Human and rat kidney tissues. ICC/IF: U87-MG cells.
  • 特記事項

    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 ab133303 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
IHC-Fr Use at an assay dependent concentration. PubMed: 21906276
Flow Cyt Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Predicted molecular weight: 67 kDa.Can be blocked with NADPH oxidase 4 peptide (ab155071).
IHC-P 1/100 - 1/600. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/500.

ターゲット情報

  • 機能
    Constitutive NADPH oxidase which generates superoxide intracellularly upon formation of a complex with CYBA/p22phox. Regulates signaling cascades probably through phosphatases inhibition. May function as an oxygen sensor regulating the KCNK3/TASK-1 potassium channel and HIF1A activity. May regulate insulin signaling cascade. May play a role in apoptosis, bone resorption and lipolysaccharide-mediated activation of NFKB. May produce superoxide in the nucleus and play a role in regulating gene expression upon cell stimulation. Isoform 3 is not functional. Isoform 4 displays an increased activity. Isoform 5 and isoform 6 display reduced activity.
  • 組織特異性
    Expressed by distal tubular cells in kidney cortex and in endothelial cells (at protein level). Widely expressed. Strongly expressed in kidney and to a lower extent in heart, adipocytes, hepatoma, endothelial cells, skeletal muscle, brain, several brain tumor cell lines and airway epithelial cells.
  • 配列類似性
    Contains 1 FAD-binding FR-type domain.
    Contains 1 ferric oxidoreductase domain.
  • 発生段階
    Expressed in fetal kidney and fetal liver.
  • 翻訳後修飾
    Isoform 3 and isoform 4 are N-glycosylated. Isoform 4 glycosylation is required for its proper function.
  • 細胞内局在
    Endoplasmic reticulum membrane. Cell membrane. Cell junction > focal adhesion. Nucleus. May localize to plasma membrane and focal adhesions. According to PubMed:15927447, may also localize to the nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Kidney oxidase 1 antibody
    • Kidney oxidase-1 antibody
    • Kidney superoxide producing NADPH oxidase antibody
    • Kidney superoxide-producing NADPH oxidase antibody
    • KOX 1 antibody
    • KOX antibody
    • Kox-1 antibody
    • KOX1 antibody
    • NADPH antibody
    • NADPH oxidase 4 antibody
    • Nox4 antibody
    • NOX4_HUMAN antibody
    • Renal NAD(P)H oxidase antibody
    • Renal NAD(P)H-oxidase antibody
    • RENOX antibody
    see all

画像

  • Immunocytochemistry/Immunofluorescence analysis of U87-MG cells labelling NADPH with purified ab133303 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 (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Flow cytometry analysis of U87-MG (human glioblastoma) cells labeling with purified ab133303 at 1/230 dilution ( 10ug/ml) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077) )(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

  • All lanes : Anti-NADPH oxidase 4 antibody [UOTR1B493] (ab133303) at 1/2000 dilution (purified)

    Lane 1 : U87-MG whole cell lysate
    Lane 2 : JAR whole cell lysate

    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: 67 kDa
    Observed band size: 67 kDa



    Blocking and dilution buffer: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling NADPH oxidase 4 with purified ab133303 at a dilution of 1/600. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling NADPH oxidase 4 with purified ab133303 at a dilution of 1/600. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • All lanes : Anti-NADPH oxidase 4 antibody [UOTR1B493] (ab133303) at 1/1000 dilution (unpurified)

    Lane 1 : Fetal kidney cell lysates
    Lane 2 : U87-MG cell lysates
    Lane 3 : 293 cell lysates
    Lane 4 : JAR cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 67 kDa

  • Immunohistochemistry (Frozen sections) analysis of rat lung tissue taken from rats with monocrotaline-exposure/pneumonectomy, labelling NOX4 with unpurified ab133303.

参考文献

This product has been referenced in:
  • Hou D  et al. Increased oxidative stress mediates the antitumor effect of PARP inhibition in ovarian cancer. Redox Biol 17:99-111 (2018). Read more (PubMed: 29684820) »
  • He Y  et al. Palmitic acid, but not high-glucose, induced myocardial apoptosis is alleviated by N-acetylcysteine due to attenuated mitochondrial-derived ROS accumulation-induced endoplasmic reticulum stress. Cell Death Dis 9:568 (2018). Read more (PubMed: 29752433) »

See all 33 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (RPE1 cells)
Gel Running Conditions
Reduced Denaturing (Bis Tris 4-12%)
Loading amount
70000 cells
Treatment
esiRNA for 72h
Specification
RPE1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

投稿 Feb 28 2018

Application
Western blot
Sample
Pig Tissue lysate - whole (Kidney)
Gel Running Conditions
Reduced Denaturing
Loading amount
35 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Abcam user community

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投稿 Nov 29 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate
Permeabilization
No
Specification
Lung
Blocking step
Dako non-serum block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 27°C
Fixative
Formaldehyde
Username

Jim Hsiao

Verified customer

投稿 Oct 31 2017

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

Dr. William Zhang

Verified customer

投稿 Sep 21 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% formalin
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Abcam user community

Verified customer

投稿 Jun 12 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

投稿 Jun 12 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
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Abcam user community

Verified customer

投稿 Jun 12 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Stomach cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Stomach cancer
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

投稿 Jun 12 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293T)
Permeabilization
Yes - 0.1% Triton X100
Specification
HEK293T
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 May 17 2017

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

Abcam user community

Verified customer

投稿 Nov 17 2016

1-10 of 13 Abreviews or Q&A

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

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