Anti-ADAM9 Antibody Picoband®

Western blot analysis of ADAM9 using anti-ADAM9 antibody (A03074-2).
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 U-87MG whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: human U20S whole cell lysates,
Lane 6: human Hela whole cell lysates,
Lane 7: human Hacat whole cell lysates,
Lane 8: human SiHa 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-ADAM9 antigen affinity purified polyclonal antibody (Catalog # A03074-2) 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 ADAM9 at approximately 75 kDa. The expected band size for ADAM9 is at 90 kDa.

Western blot analysis of ADAM9 using anti-ADAM9 antibody (A03074-2).
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 lung tissue lysates,
Lane 2: rat liver tissue lysates,
Lane 3: rat heart tissue lysates,
Lane 4: rat C6 whole cell lysates,
Lane 5: mouse lung tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse heart tissue lysates,
Lane 8: mouse C2C12 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-ADAM9 antigen affinity purified polyclonal antibody (Catalog # A03074-2) 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 ADAM9 at approximately 75 kDa. The expected band size for ADAM9 is at 90 kDa.

Flow Cytometry analysis of U251 cells using anti-ADAM9 antibody (A03074-2).
Overlay histogram showing U251 cells stained with A03074-2 (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-ADAM9 Antibody (A03074-2, 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.