Anti-IRF3 Antibody (monoclonal, 3B4)

• Catalog Number: orb738475
• Category: Antibodies
• Description: Anti-IRF3 Antibody (monoclonal, 3B4). Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Mouse, Rat.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG2a
• Conjugation: Unconjugated
• Reactivity: Mouse, Rat
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived mouse IRF3 recombinant protein (Position: M1-I419).
• UniProt ID: P70671
• MW: 50-55 kDa
• Tested applications: FC, ICC, IF, WB
• Application notes: Western blot, 0.25-0.5μg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5μg/ml, Mouse Flow Cytometry (Fixed), 1-3μg/1x106 cells, Mouse. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Antibody Type: Primary Antibody
• Clone Number: 3B4
• 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: Solute carrier family 2, facilitated glucose trans
• Note: For research use only

Flow Cytometry analysis of HEPA1-6 cells using anti-IRF3 antibody. Overlay histogram showing HEPA1-6 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-IRF3 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.

IF analysis of IRF3 using anti-IRF3 antibody. IRF3 was detected in immunocytochemical section of RM-1 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-IRF3 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of IRF3 using anti-IRF3 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: rat brain tissue lysates, Lane 2: rat C6 whole cell lysates, Lane 3: mouse lung tissue lysates, Lane 4: mouse brain tissue lysates, Lane 5: mouse NIH/3T3 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-IRF3 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 IRF3 at approximately 50-55 KD. The expected band size for IRF3 is at 50-55 KD.