Recombinant
RabMAb

Recombinant Anti-NADPH oxidase 4 antibody [UOTR1B492] - BSA and Azide free (ab230322)

Overview

  • Product name
    Anti-NADPH oxidase 4 antibody [UOTR1B492] - BSA and Azide free
    See all NADPH oxidase 4 primary antibodies
  • Description
    Rabbit monoclonal [UOTR1B492] to NADPH oxidase 4 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IP, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Rat, Dog, Human
  • Immunogen

    Synthetic peptide within Human NADPH oxidase 4 aa 500 to the C-terminus. The exact sequence is proprietary.
    (Peptide available as ab179799)

  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab230322 is a PBS-only buffer format of ab109225. Please refer to ab109225 for recommended dilutions, protocols, and image data.

     

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab230322 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-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 63 kDa (predicted molecular weight: 67 kDa).Can be blocked with NADPH oxidase 4 peptide (ab179799).
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

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

  • Flow Cytometry analysis of U87-MG (human glioblastoma) cells labeling NADPH oxidase 4 with purified ab109225 at 1/280 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

  • ab109225 (purified) at 1/30 immunoprecipitating NADPH oxidase 4 in HEK293. For western blotting, a HRP-conjugated anti-rabbit antibody was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

  • Immunofluorescent staining of HeLa cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab109225 at a dilution of 1/200. An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counter stained with DAPI. The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 594 goat anti-mouse was used at a dilution of 1/500.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

  • Immunohistochemical staining of paraffin embedded mouse cerebral cortex with purified ab109225 at a dilution of 1/500. A HRP goat anti-rabbit (ab97051) was used as the secondary antibody at a dilution of 1/500 and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

  • Immunohistochemical staining of paraffin embedded human stomach with purified ab109225 at a dilution of 1/500. A HRP goat anti-rabbit (ab97051) was used as the secondary antibody at a dilution of 1/500 and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue using unpurified ab109225.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

  • ICC/IF image of unpurified ab109255 stained HeLa 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 (ab109225, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

  • Unpurified ab109225 staining Nox4 in HeLa cells treated with (-)-cannabidiol (ab120448), by ICC/IF. Increase in Nox4 expression correlates with increased concentration of (-)-cannabidiol, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120448 ((-)-cannabidio) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab109225 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109225).

References

ab230322 has not yet been referenced specifically in any publications.

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