In biotin labeling studies, an amine-specific biotinylated agent, biotin N-hydroxyl-succinimide ester (NHS-Biotin; Sigma, USA), was dissolved in DMSO. Before adding biotin, the pH of the ACE2 protein solution was adjusted to 9 with 0.1 M carbonate buffer. 1 mg of ACE2 protein and 125 µg of biotin were combined and incubated at room temperature for 4 h. By overnight dialyzing against PBS, unreacted biotin was removed from the mixture.
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Biotin Labeling of ACE2 Protein
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Corresponding Organization :
Other organizations : Bursa Uludağ Üni̇versi̇tesi̇, Scientific and Technological Research Council of Turkey, TÜBİTAK Marmara Research Center
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Variable analysis
independent variables
- Concentration of biotin N-hydroxyl-succinimide ester (NHS-Biotin)
- Degree of biotinylation of ACE2 protein
- PH of ACE2 protein solution (adjusted to 9 with 0.1 M carbonate buffer)
- Incubation time (4 hours at room temperature)
- Removal of unreacted biotin by overnight dialysis against PBS
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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