E.coli-derived human ACE2 recombinant protein (Position: Q18-F314). Human ACE2 shares 80.5% and 80.8% amino acid (aa) sequence identity with mouse and rat ACE2, respectively.
Conjugation:
Unconjugated
Anti-ACE2 Antibody. Tested in WB, Flow Cytometry, ELISA applications. This antibody reacts with Human.
Clonality:
Polyclonal
Concentration:
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Application Notes: Western blot, 0.25-0.5 µg/ml, Human Flow Cytometry (Fixed), 1-3 µg/1x106 cells, Human ELISA, 0.1-0.5 µg/ml, -. Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml
Flow Cytometry analysis of SH-SY5Y cells using anti-ACE2 antibody. Overlay histogram showing SH-SY5Y 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-ACE2 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. 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.
Western blot analysis of ACE2 using anti-ACE2 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 Caco-2 whole cell lysates, Lane 2: human Hacat 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-ACE2 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, 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 ACE2 at approximately 120 kDa. The expected band size for ACE2 is at 93 kDa.
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