Anti-COX IV COX4I1 Antibody Picoband® (monoclonal, 4G11)

Western blot analysis of COX IV using anti-COX IV antibody (M05442-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HEPG2 whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human HEK293 whole cell lysates,
Lane 4: human T47D whole cell lysates,
Lane 5: human CACO-2 whole cell lysates,
Lane 6: human K562 whole cell lysates,
Lane 7: human Hela whole cell lysates,
Lane 8: rat brain tissue lysates,
Lane 9: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-COX IV antigen affinity purified monoclonal antibody (Catalog # M05442-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for COX IV at approximately 17KD. The expected band size for COX IV is at 17KD.

The results of western blot showed that protein expression levels of Cox4I1, Uqcr11, Atp5g1, and Atp5d were significantly lower in the Ctrl group than in the HFHS–POF group. (* p < 0.05 vs. Ctrl group; ** p < 0.01 vs. Ctrl group; t ‐test, n = 3).
Index in PubMed under a CC BY license. PMID: 40979580

IHC analysis of COX IV using anti-COX IV antibody (M05442-1).
COX IV was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COX IV Antibody (M05442-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of COX IV using anti-COX IV antibody (M05442-1).
COX IV was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COX IV Antibody (M05442-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of COX IV using anti-COX IV antibody (M05442-1).
COX IV was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COX IV Antibody (M05442-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

Flow Cytometry analysis of U937 cells using anti- COX IV antibody (M05442-1).
Overlay histogram showing U937 cells stained with M05442-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-COX IV Antibody (M05442-1, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.