Anti-TNF Receptor I/TNFRSF1A Antibody Picoband™

Figure 2. IHC analysis of TNF Receptor I using anti-TNF Receptor I antibody (A00294-3).
TNF Receptor I was detected in paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TNF Receptor I Antibody (A00294-3) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of TNF Receptor I using anti-TNF Receptor I antibody (A00294-3).
TNF Receptor I was detected in paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TNF Receptor I Antibody (A00294-3) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Figure 5. Flow Cytometry analysis of U937 cells using anti-TNF Receptor I antibody (A00294-3).
Overlay histogram showing U937 cells stained with A00294-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TNF Receptor I Antibody (A00294-3,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 4. IHC analysis of TNF Receptor I using anti-TNF Receptor I antibody (A00294-3).
TNF Receptor I was detected in paraffin-embedded section of mouse lung tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TNF Receptor I Antibody (A00294-3) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Figure 1. Western blot analysis of TNF Receptor I? using anti-TNF Receptor I? antibody (A00294-3).
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 Hela whole cell lysate,
Lane 2: human K562 whole cell lysate,
Lane 3: human Caco-2 whole cell lysate,
Lane 4: rat liver tissue lysates,
Lane 5: mouse small intestine tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse HEPA1-6 whole cell lysate.
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 rabbit anti-TNF Receptor I? antigen affinity purified polyclonal antibody (Catalog # A00294-3) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for TNF Receptor I? at approximately 50KD. The expected band size for TNF Receptor I? is at 50KD.