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Ethyl chloroformate (27 mg, 0.25 mmol) in THF (3 mL) was added with stirring.
H2O2 (30%, 7.5 mL) was slowly added, with continuous stirring, for 30 min at ambient temperature.
A peripheral blood sample (2 ml) was collected for each patient (Supplementary Methods online).
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58 U/mL was obtained.
DKK1 protein (0.5 μg/ml) was used for stimulation.
FA (200 μg/ml) was added to appropriate wells.
Limit of detection of 0.51 μg/ml was estimated.
Consequently, a detection limit of 0.4 mIU/mL was achieved.
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