Anti-TSG101 Antibody Picoband®

Western blot analysis of TSG101 using anti-TSG101 antibody (A01233-2).
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 50ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: human Colo320 whole cell lysates,
Lane 3: human Sw620 whole cell lysates,
Lane 4: human Caco-2 whole cell lysates,
Lane 5: human Hek293 whole cell lysates,
Lane 6: human K562 whole cell lysates,
Lane 7: human HepG2 whole cell lysates,
Lane 8: human THP-1 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-TSG101 antigen affinity purified polyclonal antibody (Catalog # A01233-2) 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 TSG101 at approximately 44KD. The expected band size for TSG101 is at 44KD.

Western blot analysis of TSG101 using anti-TSG101 antibody (A01233-2).
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 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat testis tissue lysates,
Lane 3: rat liver tissue lysates,
Lane 4: rat spleen tissue lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse testis tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse spleen tissue 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-TSG101 antigen affinity purified polyclonal antibody (Catalog # A01233-2) 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 TSG101 at approximately 44KD. The expected band size for TSG101 is at 44KD.

Immunoprecipitating (IP) TSG101 in HepG2 whole cell lysate.
Western blot analysis of TSG101 using anti-TSG101 antibody (A01233-2);
Lane 1: HepG2 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-TSG101 antibody in HepG2 whole cell lysate;
Lane 3: anti-TSG101 antibody (2μg) + HepG2 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-TSG101 antigen affinity purified polyclonal antibody (A01233-2) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Light Chain). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for TSG101 at approximately 44 kDa. The expected band size for TSG101 is at 44 kDa.

Flow Cytometry analysis of SiHa cells using anti-TSG101 antibody (A01233-2).
Overlay histogram showing SiHa cells stained with A01233-2 (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-TSG101 Antibody (A01233-2, 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.