Anti-ERAL1 Antibody Picoband®

Western blot analysis of ERAL1 using anti-ERAL1 antibody (A11370-1).
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 293T whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human A549 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-ERAL1 antigen affinity purified polyclonal antibody (A11370-1) 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 ERAL1 at approximately 40 kDa. The expected band size for ERAL1 is at 48 kDa.

IHC analysis of ERAL1 using anti-ERAL1 antibody (A11370-1).
ERAL1 was detected in a paraffin-embedded section of human ovarian 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-ERAL1 Antibody (A11370-1) 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.

Immunoprecipitating ERAL1 in K562 whole cell lysate.
Western blot analysis of ERAL1 using anti-ERAL1 antibody (A11370-1).
Lane 1: K562 whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-ERAL1 antibody in K562 whole cell lysate,
Lane 3: anti-ERAL1 antibody (2μg) + K562 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-ERAL1 antigen affinity purified polyclonal antibody (A11370-1) 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 ERAL1 at approximately 40 kDa. The expected band size for ERAL1 is at 48 kDa.