Anti-NADPH oxidase 4 antibody (ab79971)

Overview

  • Product name
    Anti-NADPH oxidase 4 antibody
    See all NADPH oxidase 4 primary antibodies
  • Description
    Rabbit polyclonal to NADPH oxidase 4
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Sheep, Cow, Orangutan
  • Immunogen

    Synthetic peptide derived from within residues 100 - 200 of Human NOX4.

  • Positive control
    • 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

Properties

Applications

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.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml.

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

  • 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.

  • 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.

    Secondary
    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
  • 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.

References

ab79971 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293T)
Permeabilization
Yes - 0.1% Triton X-100
Specification
HEK293T
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
Username

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Submitted Apr 25 2017

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