Flow cytometric analysis of COX IV expression in HepG2 cells using anti-COX IV antibody (Cat#69431,1:2,000). Green, isotype control; red, COX IV.

Western blotting analysis using anti-COX IV antibody (Cat#69431). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-COX IV antibody (Cat#69431,1:5,000) and HRP-conjugated goat anti rabbit secondary antibody (Cat#201,1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).

Western blotting analysis using anti-COX4I1 antibody (Cat#69431). COX4I1 expression in wild type (WT) and COX4I1 shRNA knockdown (KD) HT-1080 cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with anti-COX4I1 antibody (Cat#69431, 1:2,500) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).

Validation of COX IV knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT-1080 cells were stained with anti-COX IV antibody (Cat#69431, 1:2,000) and analyzed using BD flow cytometer.

Immunocytochemical staining of HT-1080 cells using anti-COXIV antibody (Cat#69431, 1:1,000), Top panel: wild-type (WT); Bottom panal: COXIV shRNA knockdown (KD). Nuclei were stained blue with DAPI; COXIV was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

Immunohistochemistry was performed on paraffin-embedded mouse brain using anti-COX IV antibody (Cat#69431, 1:100). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar: 25 μm.