Additional file 2.
Optimization of the measurement analysis of ACE2 activity. ACE2 activity was measured chronologically with the fluorogenic substrate and plotted for 90 min. The relative fluorescence unit (RFU) values increased within 70 minutes and then declined. Each plot is a representative result of three independent experiments. In each experiment, a rACE2 (a standard) and a reference serum (a mixture of sera from 28 healthy subjects) were assayed simultaneously with sample sera from healthy subjects, control patients, or vasculopathy patients. The difference of the ACE2 enzyme activity between healthy subjects and vasculopathy patients was statistically significant (P < 0.01) both at 60 and 70 minutes of the incubation.
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Takahashi et al. Arthritis Research & Therapy 2010 12:R85 doi:10.1186/ar3012