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Recombinant Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Flow Cytometry - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
  • Immunocytochemistry/ Immunofluorescence - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
  • Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [UOTR1B493] to NADPH oxidase 4 - BSA and Azide free
  • Suitable for: ICC/IF, IHC-P, WB, Flow Cyt
  • Reacts with: Rat, Human

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Overview

  • Product name

    Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free
    See all NADPH oxidase 4 primary antibodies
  • Description

    Rabbit monoclonal [UOTR1B493] to NADPH oxidase 4 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Rat, Human
  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab155071)

  • Positive control

    • IHC-P: Human and rat kidney tissues. ICC/IF: U87-MG cells. Flow Cyt: U87-MG
  • General notes

    Ab216654 is the carrier-free version of ab133303. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab216654 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    UOTR1B493
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Blood
    • Other
    • Signal Transduction
    • Metabolism
    • Drug metabolism
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Drug metabolism
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Alternative Versions

    • Anti-NADPH oxidase 4 antibody [UOTR1B493] (ab133303)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunizing Peptide (Blocking)

    • NADPH oxidase 4 peptide (ab155071)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • U-87 MG nuclear extract lysate (ab14903)
    • NIH/3T3 whole cell lysate (ab7179)
  • Recombinant Protein

    • Recombinant Human NADPH oxidase 4 protein (ab112414)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab216654 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
ICC/IF
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

WB
Use at an assay dependent concentration. Predicted molecular weight: 67 kDa.Can be blocked with NADPH oxidase 4 peptide (ab155071).
Flow Cyt
Use at an assay dependent concentration.
Notes
ICC/IF
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

WB
Use at an assay dependent concentration. Predicted molecular weight: 67 kDa.Can be blocked with NADPH oxidase 4 peptide (ab155071).
Flow Cyt
Use at an assay dependent concentration.

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.
  • Target information above from: UniProt accession Q9NPH5 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 50507 Human
    • Entrez Gene: 85431 Rat
    • Omim: 605261 Human
    • SwissProt: Q9NPH5 Human
    • SwissProt: Q924V1 Rat
    • Unigene: 371036 Human
    • Unigene: 14744 Rat
    • 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 - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
      Flow Cytometry - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133303).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133303).

    • Immunocytochemistry/ Immunofluorescence - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
      Immunocytochemistry/ Immunofluorescence - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)

      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).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133303).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133303).

    • Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)
      Anti-NADPH oxidase 4 antibody [UOTR1B493] - BSA and Azide free (ab216654)

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (11)

    Publishing research using ab216654? Please let us know so that we can cite the reference in this datasheet.

    ab216654 has been referenced in 11 publications.

    • Cremonini E  et al. Anthocyanins protect the gastrointestinal tract from high fat diet-induced alterations in redox signaling, barrier integrity and dysbiosis. Redox Biol 26:101269 (2019). PubMed: 31330482
    • Li XQ  et al. Liquiritin suppresses UVB-induced skin injury through prevention of inflammation, oxidative stress and apoptosis through the TLR4/MyD88/NF-?B and MAPK/caspase signaling pathways. Int J Mol Med 42:1445-1459 (2018). PubMed: 29901082
    • Hakami NY  et al. Trichostatin A, a histone deacetylase inhibitor suppresses NADPH Oxidase 4-Derived Redox Signalling and Angiogenesis. J Cell Mol Med 20:1932-44 (2016). WB ; Human . PubMed: 27297729
    • Zhao P  et al. Total Glucosides of Danggui Buxue Tang Attenuate BLM-Induced Pulmonary Fibrosis via Regulating Oxidative Stress by Inhibiting NOX4. Oxid Med Cell Longev 2015:645814 (2015). WB ; Rat . PubMed: 26347805
    • Gölz L  et al. LPS from P. gingivalis and hypoxia increases oxidative stress in periodontal ligament fibroblasts and contributes to periodontitis. Mediators Inflamm 2014:986264 (2014). WB, IHC, ICC/IF ; Human . PubMed: 25374447
    View all Publications for this product

    Customer reviews and Q&As

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