MED28 Rabbit Polyclonal Antibody

Flow Cytometry analysis of THP-1 cells using anti-MED28 antibody. Overlay histogram showing THP-1 cells (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-MED28 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Immunoprecipitating MED28 in HepG2 whole cell lysate. Western blot analysis of MED28 using anti-MED28 antibody; Lane 1: HepG2 whole cell lysates (30 ug); Lane 2: Rabbit control IgG instead of anti-MED28 antibody in HepG2 whole cell lysate; Lane 3: anti-MED28 antibody (2 µg) + HepG2 whole cell lysate (500 µg). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-MED28 antigen affinity purified polyclonal antibody at a dilution of 0.5 µg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for MED28 at approximately 24 kDa. The expected band size for MED28 is at 20 kDa.

Western blot analysis of MED28 using anti-MED28 antibody. 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 HEL whole cell lysates, Lane 2: human THP-1 whole cell lysates, Lane 3: human K562 whole cell lysates, Lane 4: human HepG2 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-MED28 antigen affinity purified polyclonal antibody 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 with Tanon 5200 system. A specific band was detected for MED28 at approximately 24 kDa. The expected band size for MED28 is at 20 kDa.