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Enzyme-linked immunosorbent assay (ELISA) assays will be carried out for the determination of serum BDNF (R&D Systems, Inc, Minneapolis, MN, USA) and total OCN (Human Osteocalcin ELISA kit KAQ1381). After incubation with 5 mg/ml hydroxylapatite (Invitrogen-Thermo Fisher Scientific, Carlsbad, CA, USA) to remove carboxylated OCN [87], serum uncarboxylated OCN will be measured by ELISA. Carboxylated OCN will be determined by subtracting uncarboxylated OCN from the total OCN [74].
JMIR Res Protoc 2023;12:e50030
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