Anti-ATF2 Antibody Picoband™

Figure 1. Western blot analysis of ATF2 using anti-ATF2 antibody (PB9131).
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 K562 whole cell lysates,
Lane 3: human SH-SY5Y whole cell lysates,
Lane 4: human U87 whole cell lysates,
Lane 5: human A549 whole cell lysates,
Lane 6: human MOLT4 whole cell lysates,
Lane 7: human HEL 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 rabbit anti-ATF2 antigen affinity purified polyclonal antibody (Catalog # PB9131) 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 1:5000 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 ATF2 at approximately 65-70 kDa. The expected band size for ATF2 is at 65-70 kDa.

Figure 2. Western blot analysis of ATF2 using anti-ATF2 antibody (PB9131).
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 thymus tissue lysates,
Lane 3: mouse brain tissue lysates,
Lane 4: mouse thymus tissue 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 rabbit anti-ATF2 antigen affinity purified polyclonal antibody (Catalog # PB9131) 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 1:5000 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 ATF2 at approximately 65-70 kDa. The expected band size for ATF2 is at 65-70 kDa.

Figure 3. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in paraffin-embedded section of human intestinal cancer 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-ATF2 Antibody (PB9131) 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 4. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in paraffin-embedded section of mouse brain 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-ATF2 Antibody (PB9131) 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. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in paraffin-embedded section of rat brain 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-ATF2 Antibody (PB9131) 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 6. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in frozen section of rat brain tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ATF2 Antibody (PB9131) 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 7. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in frozen section of mouse brain tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ATF2 Antibody (PB9131) 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 8. Flow Cytometry analysis of K562 cells using anti-ATF2 antibody (PB9131).
Overlay histogram showing K562 cells stained with PB9131 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATF2 Antibody (PB9131,1μg/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 9. Flow Cytometry analysis of HepG2 cells using anti-ATF2 antibody (PB9131).
Overlay histogram showing HepG2 cells stained with PB9131 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATF2 Antibody (PB9131,1μg/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.