Recombinant
RabMAb

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

Overview

  • Product name
    Anti-NADPH oxidase 4 antibody [UOTR1B493]
    See all NADPH oxidase 4 primary antibodies
  • Description
    Rabbit monoclonal [UOTR1B493] to NADPH oxidase 4
  • Tested applications
    Suitable for: IHC-Fr, Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human NADPH oxidase 4. The exact sequence is proprietary.
    (Peptide available as ab155071)

  • Positive control
    • WB: Human fetal kidney tissue lysate, U87-MG, 293, C6, PC-12, NIH 3T3 and JAR cell lysates. IHC-P: Human, mouse and rat kidney tissues. ICC/IF: U87-MG cells.
  • General notes

    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.

Properties

Applications

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.

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

ICC/IF 1/500.

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    Contains 1 FAD-binding FR-type domain.
    Contains 1 ferric oxidoreductase domain.
  • Developmental stage
    Expressed in fetal kidney and fetal liver.
  • Post-translational
    modifications
    Isoform 3 and isoform 4 are N-glycosylated. Isoform 4 glycosylation is required for its proper function.
  • Cellular localization
    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
  • Database links
  • Alternative names
    • 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

Images

  • 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse 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 1/2000 dilution (purified)

    Lane 1 : C6 whole cell lysate
    Lane 2 : PC-12 whole cell lysate
    Lane 3 : NIH/3T3 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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.

  • 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
    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
    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.

References

This product has been referenced in:
  • Wang M  et al. TRB3 mediates advanced glycation end product-induced apoptosis of pancreatic ß-cells through the protein kinase C ß pathway. Int J Mol Med 40:130-136 (2017). WB ; Rat . Read more (PubMed: 28534945) »
  • Ju HQ  et al. Mutant Kras- and p16-regulated NOX4 activation overcomes metabolic checkpoints in development of pancreatic ductal adenocarcinoma. Nat Commun 8:14437 (2017). WB . Read more (PubMed: 28232723) »

See all 20 Publications for this product

Customer reviews and Q&As

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
Username

Abcam user community

Verified customer

Submitted 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

Submitted 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

Submitted 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
Username

Abcam user community

Verified customer

Submitted 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

Submitted 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
Username

Abcam user community

Verified customer

Submitted 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

Submitted 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

Submitted 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

Submitted Nov 17 2016

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

Seong Hoon Kim

Verified customer

Submitted Jun 23 2016

1-10 of 12 Abreviews or Q&A

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