Anti-Interferon gamma/IFNG Antibody Picoband™ test

Figure 3. IHC analysis of IFNG using anti-IFNG antibody (A00393-3).

IFNG was detected in paraffin-embedded section of human sarcoma 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-IFNG Antibody (A00393-3) 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 2. Western blot analysis of IFNG using anti-IFNG antibody (A00393-3).

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: rat brain tissue lysates,
• Lane 2: rat lung tissue lysates,
• Lane 3: mouse brain tissue lysates,
• Lane 4: mouse ovary tissue lysates,
• Lane 5: mouse HEPA1-6 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 rabbit anti-IFNG antigen affinity purified polyclonal antibody (Catalog # A00393-3) 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 IFNG at approximately 19KD. The expected band size for IFNG is at 19KD.

Figure 1. Western blot analysis of IFNG using anti-IFNG antibody (A00393-3).

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 lysates,
• Lane 2: human placenta tissue lysates,
• Lane 3: human U-87MG whole cell lysates,
• Lane 4: human HL-60 whole cell lysates,
• Lane 5: human U-937 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 rabbit anti-IFNG antigen affinity purified polyclonal antibody (Catalog # A00393-3) 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 IFNG at approximately 19KD. The expected band size for IFNG is at 19KD.

Figure 6. IHC analysis of IFNG using anti-IFNG antibody (A00393-3).

IFNG was detected in paraffin-embedded section of human tonsil 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-IFNG Antibody (A00393-3) 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.