Anti-Cyt 19/As3mt Antibody Picoband®

Western blot analysis of Cyt 19/As3mt using anti-Cyt 19/As3mt antibody (A05107-2).
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 REH whole cell lysates,
Lane 2: human SH-SY5Y whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: rat liver tissue lysates,
Lane 5: rat heart tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse heart 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-Cyt 19/As3mt antigen affinity purified polyclonal antibody (A05107-2) 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 Cyt 19/As3mt at approximately 42-45 kDa. The expected band size for Cyt 19/As3mt is at 42 kDa.

IHC analysis of Cyt 19/As3mt using anti-Cyt 19/As3mt antibody (A05107-2).
Cyt 19/As3mt was detected in a paraffin-embedded section of human renal 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 2 μg/ml rabbit anti-Cyt 19/As3mt Antibody (A05107-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of Cyt 19/As3mt using anti-Cyt 19/As3mt antibody (A05107-2).
Cyt 19/As3mt was detected in an immunocytochemical section of A549 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-Cyt 19/As3mt Antibody (A05107-2) overnight at 4°C. Fluoro488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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.

Immunoprecipitating (IP) Cyt 19/As3mt in HepG2 whole cell lysate.
Western blot analysis of Cyt 19/As3mt using anti-Cyt 19/As3mt antibody (A05107-2);
Lane 1: HepG2 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-Cyt 19/As3mt antibody in HepG2 whole cell lysate;
Lane 3: anti-Cyt 19/As3mt antibody (2μg) + HepG2 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-Cyt 19/As3mt antigen affinity purified polyclonal antibody (A05107-2) 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 # AR1196-200). A specific band was detected for Cyt 19/As3mt at approximately 42 kDa. The expected band size for Cyt 19/As3mt is at 42 kDa.