Anti-Glutathione Peroxidase 4/GPX4 Antibody (monoclonal, 6I4E7)

• Catalog Number: orb1184745
• Category: Antibodies
• Description: Anti-Glutathione Peroxidase 4/GPX4 Antibody (monoclonal, 6I4E7). Tested in Flow Cytometry, WB applications. This antibody reacts with Human.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG2b
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived human Glutathione Peroxidase 4/GPX4 recombinant protein (Position: A30-F197).
• UniProt ID: P36969
• MW: 19 kDa
• Tested applications: FC, WB
• Application notes: Western blot, 0.25-0.5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Antibody Type: Primary Antibody
• Clone Number: 6I4E7
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Desmin; DES
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of Hela cells using anti-Glutathione Peroxidase 4/GPX4 antibody. Overlay histogram showing Hela cells (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-Glutathione Peroxidase 4/GPX4 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of Glutathione Peroxidase 4/GPX4 using anti-Glutathione Peroxidase 4/GPX4 antibody. 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 30 ug of sample under reducing conditions. Lane 1: human HepG2 whole cell lysates, Lane 2: human CACO-2 whole cell lysates, Lane 3: human 293T whole cell lysates, Lane 4: human K562 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-Glutathione Peroxidase 4/GPX4 antigen affinity purified monoclonal antibody 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 with Tanon 5200 system. A specific band was detected for Glutathione Peroxidase 4/GPX4 at approximately 19 kDa. The expected band size for Glutathione Peroxidase 4/GPX4 is at 22 kDa.