Anti-eIF4A2 Antibody Picoband®

Western blot analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758).
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 U251 whole cell lysates,
Lane 3: human SiHa whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat lung tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse lung 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-eIF4A2 antigen affinity purified polyclonal antibody (Catalog # PB9758) 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 eIF4A2 at approximately 46 kDa. The expected band size for eIF4A2 is at 46 kDa.

IHC analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758).
eIF4A2 was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-eIF4A2 Antibody (PB9758) 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.

IHC analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758).
eIF4A2 was detected in a paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-eIF4A2 Antibody (PB9758) 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.

IHC analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758).
eIF4A2 was detected in a paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-eIF4A2 Antibody (PB9758) 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.

IF analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758).
eIF4A2 was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-eIF4A2 Antibody (PB9758) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of SiHa cells using anti-eIF4A2 antibody (PB9758).
Overlay histogram showing SiHa cells stained with PB9758 (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-eIF4A2 Antibody (PB9758,1μg/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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758).
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 U251 whole cell lysates,
Lane 3: human SiHa whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat lung tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse lung 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-eIF4A2 antigen affinity purified polyclonal antibody (PB9758) 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-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for eIF4A2 at approximately 46 kDa. The expected band size for eIF4A2 is at 46 kDa.

Immunoprecipitating (IP) eIF4A2 in K562 whole cell lysate.
Western blot analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758);
Lane 1: K562 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-eIF4A2 antibody in K562 whole cell lysate;
Lane 3: anti-eIF4A2 antibody (2μg) + K562 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-eIF4A2 antigen affinity purified polyclonal antibody (PB9758) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BM2007). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for eIF4A2 at approximately 46 kDa. The expected band size for eIF4A2 is at 46 kDa.