Anti-PSMC6 Antibody Picoband®

Western blot analysis of PSMC6 using anti-PSMC6 antibody (A09249-2).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat testis tissue lysates,
Lane 4: rat liver tissue lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse testis tissue lysates,
Lane 7: mouse liver 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-PSMC6 antigen affinity purified polyclonal antibody (A09249-2) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for PSMC6 at approximately 42 kDa. The expected band size for PSMC6 is at 44 kDa.

IF analysis of PSMC6 using anti-PSMC6 antibody (A09249-2) and anti-Alpha Tubulin antibody (M03989-3).
PSMC6 was detected in an immunocytochemical section of U2OS cells. 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 5 μg/mL rabbit anti-PSMC6 Antibody (A09249-2) and mouse anti-Alpha Tubulin antibody (M03989-3) overnight at 4°C. Fluoro488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating PSMC6 in Hela whole cell lysate.
Western blot analysis of PSMC6 using anti-PSMC6 antibody (A09249-2).
Lane 1: Hela whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-PSMC6 antibody in Hela whole cell lysate,
Lane 3: anti-PSMC6 antibody (2μg) + Hela whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PSMC6 antigen affinity purified polyclonal antibody (A09249-2) at a dilution of 0.5 μg/mL and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Catalog # BM2007). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for PSMC6 at approximately 42 kDa. The expected band size for PSMC6 is at 44 kDa.

Flow Cytometry analysis of HeLa cells using anti-PSMC6 antibody (A09249-2).
Overlay histogram showing HeLa cells stained with A09249-2 (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-PSMC6 Antibody (A09249-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. Fluoro488 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.