Key features and details
- Mouse monoclonal [54.1] to NOX2/gp91phox
- Suitable for: WB, ICC/IF, IHC-P
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-NOX2/gp91phox antibody [54.1]
See all NOX2/gp91phox primary antibodies
DescriptionMouse monoclonal [54.1] to NOX2/gp91phox
SpecificityThis antibody is specific for NOX2/gp91phox 382-PKIAVDGP-389.
Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Human
Tissue/ cell preparation (Human): Partially Purified human neutrophil flavocytochrome b (heparin Ultrogel/lectin affinity purification in Triton X-100).
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.1% Sodium azide
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab80897 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionCritical 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 diseaseDefects 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 similaritiesContains 1 FAD-binding FR-type domain.
Contains 1 ferric oxidoreductase domain.
- Information by UniProt
- AMCBX2 antibody
- CGD antibody
- CGD91-phox antibody
All lanes : Anti-NOX2/gp91phox antibody [54.1] (ab80897) at 1 µg/ml
Lane 1 : Human liver tissue lysate - total protein (ab29889)
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : Human lymph node tissue lysate - total protein (ab29871)
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 65 kDa
Observed band size: 65 kDa
Additional bands at: 29 kDa, 35 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 12 minutes
ICC/IF image of ab80897 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab80897, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.
IHC image of ab80897 staining in normal human skin 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 ab80897, 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.
ab80897 has been referenced in 6 publications.
- Marone IM et al. TRPA1/NOX in the soma of trigeminal ganglion neurons mediates migraine-related pain of glyceryl trinitrate in mice. Brain N/A:N/A (2018). PubMed: 29985973
- Dingjan I et al. Lipid peroxidation causes endosomal antigen release for cross-presentation. Sci Rep 6:22064 (2016). PubMed: 26907999
- Wilson C et al. Contribution of NADPH oxidase to the establishment of hippocampal neuronal polarity in culture. J Cell Sci 128:2989-95 (2015). Mouse, Rat . PubMed: 26101350
- Selemidis S et al. Nox1 oxidase suppresses influenza a virus-induced lung inflammation and oxidative stress. PLoS One 8:e60792 (2013). IHC-P ; Mouse . PubMed: 23577160
- Baniulis D et al. Evaluation of two anti-gp91phox antibodies as immunoprobes for Nox family proteins: mAb 54.1 recognizes recombinant full-length Nox2, Nox3 and the C-terminal domains of Nox1-4 and cross-reacts with GRP 58. Biochim Biophys Acta 1752:186-96 (2005). PubMed: 16140048
- Burritt JB et al. Topological mapping of neutrophil cytochrome b epitopes with phage-display libraries. J Biol Chem 270:16974-80 (1995). PubMed: 7622517