NADPH Oxidase 4 Antibody

Western blot analysis of extracts from Hela using NADPH Oxidase 4 antibody. Proteins were transferred to a PVDF membrane and blocked with 5% nonfat powdered milk in PBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (GAPDH, 1/3000) was diluted with 5% nonfat powdered milk in PBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/8,000) was incubated for 45min at room temperature.

Immunohistochemical analysis of paraffin-embedded human Lung squamous cell carcinoma tissue using NADPH Oxidase 4 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80762, 1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue using NADPH Oxidase 4 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80762, 1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human ovarian carcinoma (sample 1) tissue using NADPH Oxidase 4 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P80762, 1:100 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with TSA520 . The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human ovarian carcinoma (sample 1) tissue using NADPH Oxidase 4 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P80762, 1:100 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with TSA520 . The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.