Recombinant
RabMAb

Recombinant Anti-NADPH oxidase 4 antibody [UOTR1B492] (HRP) (ab195524)

Overview

  • Product name
    Anti-NADPH oxidase 4 antibody [UOTR1B492] (HRP)
    See all NADPH oxidase 4 primary antibodies
  • Description
    Rabbit monoclonal [UOTR1B492] to NADPH oxidase 4 (HRP)
  • Host species
    Rabbit
  • Conjugation
    HRP
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human NADPH oxidase 4 aa 500 to the C-terminus.
    (Peptide available as ab179799)

  • Positive control
    • WB: HEK293 (Human embryonic kidney cell line) Whole Cell lysates. IHC-P: FFPE human normal kidney tissue sections.
  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab195524 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 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 63 kDa (predicted molecular weight: 67 kDa).Can be blocked with NADPH oxidase 4 peptide (ab179799).

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

  • Anti-NADPH oxidase 4 antibody [UOTR1B492] (HRP) (ab195524) at 1/1000 dilution + HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 67 kDa
    Observed band size: 63 kDa
    why is the actual band size different from the predicted?


    Exposure time: 90 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195524 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • IHC image of NADPH oxidase 4 staining in a section of formalin-fixed paraffin-embedded human normal kidney*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab195524 at a dilution of 1/500. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

References

This product has been referenced in:
  • Chen J  et al. Angiotensin-converting enzyme 2 priming enhances the function of endothelial progenitor cells and their therapeutic efficacy. Hypertension 61:681-9 (2013). WB ; Mouse . Read more (PubMed: 23266545) »
See 1 Publication for this product

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