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Anti-NOX4 Antibody UOTR1B492
Also for NOX4 (NM_001143836)
|A synthetic peptide corresponding to residues in human NOX4 was used as an immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||FC: 1:1000; WB: 1:1000 - 1:10000; IP: 1:10 - 1:100; IHC-P: 1:100 - 1:250; ICC: 1:100 - 1:250
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Homo sapiens NADPH oxidase 4 (NOX4), transcript variant 2|
|KOX; KOX-1; RENOX|
|NOX4 is a member of the NOX-family of enzymes that functions as the catalytic subunit of the NADPH oxidase complex. It is localized to non-phagocytic cells, where it acts as an oxygen sensor and catalyzes the reduction of molecular oxygen to various reactive oxygen species (ROS). The ROS generated by NOX4 have been implicated in numerous biological functions including signal transduction, cell differentiation and tumor cell growth.|
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Western blot - NOX4 antibody [UOTR1B492]; All lanes : Anti-NADPH oxidase 4 [UOTR1B492] antibody at 1/1000 dilution.Lane 1 : Fetal kidney lysate.Lane 2 : U87-MG lysate.Lane 3 : 293T lysate.Lane 4 : JAR lysates .Lysates/proteins at 10 µg per lane.Secondary.Standard HRP labelled goat anti-rabbit at 1/2000 dilution.Predicted band size : 67 kDa.Observed band size : 63 kDa .
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - NOX4 antibody [UOTR1B492]; Immunohistochemical analysis of paraffin-embedded Human kidney tissue using ab109225.
Immunocytochemistry/ Immunofluorescence - Anti-NOX4 antibody [UOTR1B492]; ICC/IF image of 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 overnight at +4°C. The secondary antibody (green) was , DyLight? 488 goat anti-rabbit 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.
Immunocytochemistry/ Immunofluorescence - Anti-NOX4 antibody [UOTR1B492]; ab109225 staining Nox4 in HeLa cells treated with (-)-cannabidiol , by ICC/IF. Increase in Nox4 expression correlates with increased concentration of (-)-cannabidiol, as described in literature.The cells were incubated at 37°C for 6h in media containing different concentrations of ((-)-cannabidio) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab109225 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody at 1/250 dilution was used as the secondary antibody.
Flow Cytometry - Anti-NADPH oxidase 4 antibody [UOTR1B492]; Overlay histogram showing HEK293 cells stained with ab109225 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.