Anti-NADPH oxidase 4 antibody [UOTR1B492]

• Supplier: Abcam
• Catalog: ab109225
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: UOTR1B492
• Applications: WB IP IHC-P ICC/IF
• Species Reactivities: Mouse, Rat, Dog, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human NADPH oxidase 4 aa 500-600
• Blocking Peptide: NADPH oxidase 4 peptide
• Description: Rabbit Monoclonal
• Gene: NOX4
• Conjugate: Unconjugated

Product images

Anti-NADPH oxidase 4 antibody [UOTR1B492] (ab109225) at 1/10000 dilution (purified) + JAR cell lysate at 10 µgSecondaryHRP goat anti-rabbit (H+L) at 1/1000 dilution

Immunohistochemical staining of paraffin embedded human stomach with purified ab109225 at a dilution of 1/500. A HRP goat anti-rabbit (ab97051) was used as the secondary antibody at a dilution of 1/500 and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

Immunofluorescent staining of HeLa cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab109225 at a dilution of 1/200. An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counter stained with DAPI. The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 594 goat anti-mouse was used at a dilution of 1/500.

Anti-NADPH oxidase 4 antibody [UOTR1B492] (ab109225) at 1/2000 dilution (purified) + Human fetal kidney at 10 µgSecondaryHRP anti-rabbit, specific to the non reducded form of IgG at 1/1000 dilution

ab109225 (purified) at 1/30 immunoprecipitating NADPH oxidase 4 in HEK293. For western blotting, a HRP-conjugated anti-rabbit antibody was used as the secondary antibody (1/1000).Blocking buffer and concentration: 5% NFDM/TBST.Diluting buffer and concentration: 5% NFDM /TBST.

All lanes : Anti-NADPH oxidase 4 antibody [UOTR1B492] (ab109225) at 1/1000 dilution (unpurified)Lane 1 : Fetal kidney lysateLane 2 : U87-MG lysateLane 3 : 293T lysateLane 4 : JAR lysates Lysates/proteins at 10 µg per lane.SecondaryStandard HRP labelled goat anti-rabbit at 1/2000 dilution

Immunohistochemical analysis of paraffin-embedded Human kidney tissue using unpurified ab109225.

ICC/IF image of unpurified ab109255 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 (ab109225, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, 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.

Unpurified ab109225 staining Nox4 in HeLa cells treated with (-)-cannabidiol (ab120448), 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 ab120448 ((-)-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 (ab96899) at 1/250 dilution was used as the secondary antibody.

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