Overview

  • Product name

    Anti-NOX2/gp91phox antibody
    See all NOX2/gp91phox primary antibodies
  • Description

    Rabbit polyclonal to NOX2/gp91phox
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-Fr, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rabbit, Cow, Dog
  • Immunogen

    Synthetic peptide corresponding to Human NOX2/gp91phox aa 511-560 (C terminal).
    Sequence:

    YGRPNWDNEFKTIASQHPNTRIGVFLCGPEALAETLSKQSISNSESGPRG


    (Peptide available as ab111671)

  • Positive control

    • MCF7 cell lysate. THP-1 cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab80508 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 a concentration of 1 µg/ml.
Flow Cyt Use at an assay dependent concentration.
IHC-Fr Use a concentration of 5 µg/ml.
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).

Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

As published in databases Human Protein Atlas, GeneCard and NCBI CYBB expression in heart is cell type specific therefore, we recommend including positive controls when using this antibody with heart samples.

Target

  • Function

    Critical component of the membrane-bound oxidase of phagocytes that generates superoxide. It is the terminal component of a respiratory chain that transfers single electrons from cytoplasmic NADPH across the plasma membrane to molecular oxygen on the exterior. Also functions as a voltage-gated proton channel that mediates the H(+) currents of resting phagocytes. It participates in the regulation of cellular pH and is blocked by zinc.
  • Involvement in disease

    Defects in CYBB are a cause of chronic granulomatous disease X-linked (XCGD) [MIM:306400]. Chronic granulomatous disease is a genetically heterogeneous disorder characterized by the inability of neutrophils and phagocytes to kill microbes that they have ingested. Patients suffer from life-threatening bacterial/fungal infections.
  • Sequence similarities

    Contains 1 FAD-binding FR-type domain.
    Contains 1 ferric oxidoreductase domain.
  • Post-translational
    modifications

    Glycosylated.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • AMCBX2 antibody
    • CGD antibody
    • CGD91-phox antibody
    • CY24B_HUMAN antibody
    • CYBB antibody
    • Cytochrome b 245, beta polypeptide antibody
    • Cytochrome b(558) beta chain antibody
    • Cytochrome b(558) subunit beta antibody
    • Cytochrome b-245 heavy chain antibody
    • Cytochrome b558 subunit beta antibody
    • GP91 PHOX antibody
    • gp91-1 antibody
    • gp91-phox antibody
    • GP91PHOX antibody
    • Heme-binding membrane glycoprotein gp91phox antibody
    • NADPH oxidase 2 antibody
    • Neutrophil cytochrome b 91 kDa polypeptide antibody
    • NOX2 antibody
    • p22 phagocyte B-cytochrome antibody
    • P91 PHOX antibody
    • p91-PHOX antibody
    • Superoxide-generating NADPH oxidase heavy chain subunit antibody
    see all

Images

  • Anti-NOX2/gp91phox antibody (ab80508) at 1 µg/ml + MCF7 cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 65 kDa
    Observed band size: 65 kDa
    Additional bands at: 36 kDa. We are unsure as to the identity of these extra bands.

  • ab80508 staining NOX2/gp91phox in Mouse lung tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% milk for 30 minutes at 37°C; antigen retrieval was by heat mediation with a Tris Borate EDTA buffer (pH 8.4). Samples were incubated with primary antibody (1/200 in antibody diluent) for 1 hour at 37°C. A Biotin-conjugated Goat anti-rabbit polyclonal (1/700) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab80508 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 (ab80508, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • ab80508 staining NOX2/gp91phox in Human monocyte-derivive dendritic cells by Flow Cytometry. Cells were fixed with formaldehyde and permeabilized with methanol. The sample was incubated with the primary antibody (1/200) for 30 minutes. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal (1/300) was used as the secondary antibody

    See Abreview

  • All lanes : Anti-NOX2/gp91phox antibody (ab80508) at 1/1000 dilution

    All lanes : Mouse liver whole cell lysate

    Lysates/proteins at 100 µg per lane.

    Secondary
    All lanes : HRP-conjugated Goat polyclonal to rabbit IgG at 1/5000 dilution

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Predicted band size: 65 kDa
    Observed band size: 65 kDa


    Exposure time: 5 seconds


    Primary incubated for 9 hours at 4°C.

    See Abreview

References

This product has been referenced in:

  • Tanaka Y  et al. Adiponectin promotes muscle regeneration through binding to T-cadherin. Sci Rep 9:16 (2019). Read more (PubMed: 30626897) »
  • Hu L  et al. Melatonin decreases M1 polarization via attenuating mitochondrial oxidative damage depending on UCP2 pathway in prorenin-treated microglia. PLoS One 14:e0212138 (2019). Read more (PubMed: 30742657) »
See all 39 Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews

Application
Western blot
Sample
Mouse Tissue lysate - whole (retina)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
retina
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Apr 08 2019

Application
Western blot
Sample
Rat Tissue lysate - whole (muscle)
Gel Running Conditions
Reduced Denaturing (9%)
Loading amount
60 µg
Specification
muscle
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Mr. Baptiste Jude

Verified customer

Submitted Jul 18 2016

Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 02 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 02 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Loading amount
50 µg
Specification
Skeletal muscle
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. G.K Sakellariou

Verified customer

Submitted Sep 28 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Single isolated FDB skeletal muscle fibres)
Specification
Single isolated FDB skeletal muscle fibres
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100 (for 10 hours)
Blocking step
BSA as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C

Dr. G.K Sakellariou

Verified customer

Submitted Sep 25 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lung)
Specification
Lung
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris Borate EDTA ph8.4 - 40 min.
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 37°C

Dr. Alain Fautrel

Verified customer

Submitted Apr 11 2011

Application
Western blot
Sample
Mouse Cell lysate - whole cell (skeletal muscle)
Loading amount
200 µg
Specification
skeletal muscle
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 31 2011

Application
Flow Cytometry
Sample
Human Cell (monocyte-derived dendritic cell)
Specification
monocyte-derived dendritic cell
Fixation
Formaldehyde
Permeabilization
Yes - methanol

Abcam user community

Verified customer

Submitted Oct 22 2010

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Liver)
Loading amount
100 µg
Specification
Liver
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 18 2009

1-10 of 11 Abreviews

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