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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.|
|Mouse, Rat, Dog, Human
||Lot dependent; please refer to CoA along with shipment
|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
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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 ug 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 TA307575.
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 1.43uM.
Immunocytochemistry/ Immunofluorescence - Anti-NOX4 antibody [UOTR1B492]; TA307575 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 TA307575 (5 ug/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight488 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 TA307575 (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) (0.1ug/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.