Anti-UCHL3 Antibody Picoband®

Western blot analysis of UCHL3 using anti-UCHL3 antibody (A05004-1).
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: human Jurkat whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: human A431 whole cell lysates,
Lane 5: rat spleen tissue lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse spleen tissue lysates,
Lane 8: mouse brain 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-UCHL3 antigen affinity purified polyclonal antibody (A05004-1) at 1:1000 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 UCHL3 at approximately 26 kDa. The expected band size for UCHL3 is at 26 kDa.

Western blot analysis of UCHL3 using anti-UCHL3 antibody (A05004-1).
Electrophoresis was performed on a 12% 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: human Jurkat whole cell lysates,
Lane 3: human SW620 whole cell lysates,
Lane 4: human U-87MG whole cell lysates,
Lane 5: rat testis tissue lysates,
Lane 6: mouse testis 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-UCHL3 antigen affinity purified polyclonal antibody (A05004-1) 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 UCHL3 at approximately 26 kDa. The expected band size for UCHL3 is at 26 kDa.

IF analysis of UCHL3 using anti-UCHL3 antibody (A05004-1) and anti-Alpha Tubulin antibody (M03989-3).
UCHL3 was detected in an immunocytochemical section of Hela 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-UCHL3 Antibody (A05004-1) and mouse anti-Alpha Tubulin antibody (M03989-3) overnight at 4°C. DyLight®488 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 UCHL3 in Hela whole cell lysate.
Western blot analysis of UCHL3 using anti-UCHL3 antibody (A05004-1).
Lane 1: Hela whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-UCHL3 antibody in Hela whole cell lysate,
Lane 3: anti-UCHL3 antibody (2μg) + Hela whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-UCHL3 antigen affinity purified polyclonal antibody (A05004-1) 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 UCHL3 at approximately 26 kDa. The expected band size for UCHL3 is at 26 kDa.