Product nameAnti-NADPH oxidase 4 antibody
See all NADPH oxidase 4 primary antibodies
DescriptionRabbit polyclonal to NADPH oxidase 4
Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Human
Predicted to work with: Sheep, Cow, Orangutan
Synthetic peptide derived from within residues 100 - 200 of Human NOX4.
- This antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; HepG2. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal kidney. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: A431
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab79971 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa).Can be blocked with Recombinant Human NADPH oxidase 4 protein (ab112414).|
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
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 (ab79971) at 1 µg/ml
Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 67 kDa
Observed band size: 67 kDa
Additional bands at: 48 kDa, 62 kDa (possible isoform), 94 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
ICC/IF image of ab79971 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab79971 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of NADPH oxidase 4 staining in Human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab79971, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.