Carbonate Ion → Biotin

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Following the above-mentioned method, labeled antibodies were produced using monoclonal antibodies. Anti-α-subunit monoclonal antibody (IM0365) (5 μg/mL) dissolved in 0.1 M carbonate buffer (pH 8.5) and NHS-LC-BIOTIN (PIERCE, 25 μg/mL) were mixed and stirred at room temperature for four hours using a stirrer. IgG and NHS-LC-BIOTIN were mixed at a mole ratio of 1:60. After stiring, this solution was dialyzed against physiological phosphate buffered saline (PBS) to obtain biotin-labeled antibodies.

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