IRF3 Antibody – Mouse Monoclonal

Figure 1. Western blot analysis of IRF3 using anti-IRF3 antibody (MBS1753961).
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 30ug 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 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- IRF3 antigen affinity purified monoclonal antibody (Catalog # MBS1753961) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176445) with Tanon 5200 system. A specific band was detected for IRF3 at approximately 50-55KD. The expected band size for IRF3 is at 50-55KD.

Figure 2. IF analysis of IRF3 using anti- IRF3 antibody (MBS1753961).
IRF3 was detected in immunocytochemical section of RM-1 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- IRF3 Antibody (MBS1753961) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 3. Flow Cytometry analysis of HEPA1-6 cells using anti-IRF3 antibody (MBS1753961).
Overlay histogram showing HEPA1-6 cells stained with MBS1753961 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- IRF3 Antibody (MBS1753961, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.