Anti-RPL27 Antibody Picoband®

Figure 1. Western blot analysis of RPL27 using anti-RPL27 antibody (A09736-2).
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 THP-1 whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human Caco-2 whole cell lysates,
Lane 5: human RT4 whole cell lysates,
Lane 6: human Hela whole cell lysates,
Lane 8: human K562 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-RPL27 antigen affinity purified polyclonal antibody (Catalog # A09736-2) at 0.25 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 RPL27 at approximately 16 kDa. The expected band size for RPL27 is at 16 kDa.

Figure 2. Western blot analysis of RPL27 using anti-RPL27 antibody (A09736-2).
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 liver tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat thymus tissue lysates,
Lane 4: rat RH35 whole cell lysates,
Lane 5: mouse liver tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse thymus tissue lysates,
Lane 8: 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 rabbit anti-RPL27 antigen affinity purified polyclonal antibody (Catalog # A09736-2) 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 RPL27 at approximately 16 kDa. The expected band size for RPL27 is at 16 kDa.

Figure 3. Flow Cytometry analysis of CACO-2 cells using anti-RPL27 antibody (A09736-2).
Overlay histogram showing CACO-2 cells stained with A09736-2 (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 rabbit anti-RPL27 Antibody (A09736-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Figure 4. Flow Cytometry analysis of NIH/3T3 cells using anti-RPL27 antibody (A09736-2).
Overlay histogram showing NIH/3T3 cells stained with A09736-2 (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 rabbit anti-RPL27 Antibody (A09736-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Figure 5. Flow Cytometry analysis of RH35 cells using anti-RPL27 antibody (A09736-2).
Overlay histogram showing RH35 cells stained with A09736-2 (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 rabbit anti-RPL27 Antibody (A09736-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Figure 6. Immunoprecipitating RPL27 in Hela whole cell lysate .
Western blot analysis of RPL27 using anti-RPL27 antibody (A09736-2).
Lane 1: Hela whole cell lysates (30ug)
Lane 2: Rabbit control IgG instead of anti-RPL27 antibody in Hela whole cell lysate.
Lane 3: anti-RPL27 antibody (2μg) + Hela whole cell lysate (500μg)
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-RPL27 antigen affinity purified polyclonal antibody (A09736-2) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for RPL27 at approximately 16 kDa. The expected band size for RPL27 is at 16 kDa.