Product nameAnti-NADPH oxidase 4 antibody [UOTR1B493]
See all NADPH oxidase 4 primary antibodies
DescriptionRabbit monoclonal [UOTR1B493] to NADPH oxidase 4
Tested applicationsSuitable for: IHC-Fr, Flow Cyt, WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Rat, Human
- 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. FC: U87-MG
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
PurityProtein A purified
Immunizing Peptide (Blocking)
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.
|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.|
FunctionConstitutive 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 specificityExpressed 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 similaritiesContains 1 FAD-binding FR-type domain.
Contains 1 ferric oxidoreductase domain.
Developmental stageExpressed in fetal kidney and fetal liver.
modificationsIsoform 3 and isoform 4 are N-glycosylated. Isoform 4 glycosylation is required for its proper function.
Cellular localizationEndoplasmic 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
- Kidney oxidase 1 antibody
- Kidney oxidase-1 antibody
- Kidney superoxide producing NADPH oxidase antibody
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.
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 67 kDa
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.
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).
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.
Paraffin-embedded human normal colon and colorectal cancer tissue stained for NADPH oxidase 4 using ab133303 at 1/100 dilution in immunohistochemical analysis.
NOX4 expression in CRC samples compared with corresponding normal tissue. (a) Normal tissue; (b) CRC, scored as (−); (c) CRC, scored as (+); (d) CRC, scored as (++); (e) CRC, scored as (+++) (scale bar, 100 μm).
CRC = colorectal cancer.
From Figure 1D of Lin et al.
Reproduced under the Creative Commons Attribution License (CC-BY).
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 1/2000 dilution (purified)
Lane 1 : U87-MG whole cell lysate
Lane 2 : JAR whole cell lysate
Lysates/proteins at 20 µg per lane.
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 (Frozen sections) analysis of rat lung tissue taken from rats with monocrotaline-exposure/pneumonectomy, labelling NOX4 with unpurified ab133303.
This product has been referenced in:
- Oglio R et al. Participation of NADPH 4 oxidase in thyroid regulation. Mol Cell Endocrinol 480:65-73 (2019). Read more (PubMed: 30316800) »
- Witten ML et al. Early life inhalation exposure to mine tailings dust affects lung development. Toxicol Appl Pharmacol 365:124-132 (2019). Read more (PubMed: 30641074) »