alpha 2a Adrenergic Receptor/A

Flow Cytometry analysis of HEL cells using anti-alpha 2a Adrenergic Receptor/ADRA2A antibody. Overlay histogram showing HEL cells (Blue line). The cells were fixed with 4% Fixative and blocked with 10% normal goat serum. And then incubated with rabbit anti-alpha 2a Adrenergic Receptor/ADRA2A 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of alpha 2a Adrenergic Receptor/ADRA2A using anti-alpha 2a Adrenergic Receptor/ADRA2A 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: rat pancreas tissue lysates, Lane 2: rat small intestine tissue lysates, Lane 3: mouse pancreas tissue lysates, Lane 4: mouse small intestine tissue lysate. 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-alpha 2a Adrenergic Receptor/ADRA2A 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 alpha 2a Adrenergic Receptor/ADRA2A at approximately 55 kDa. The expected band size for alpha 2a Adrenergic Receptor/ADRA2A is at 49 kDa.