反应详情

STI is dissolved in 50 ml of borate-saline buffer pH 8.5, to a concentration of 16 mg/ml (38 μmole). N-hydroxysuccinimide ester of N-acetyl thyroxine (378 μmole, 346 mg) is dissolved in 50 ml of DMF in a separate container. Aliquots (2 ml) of the active ester in DMF are added to the STI at 5 minute intervals until all the active ester has been added. The reaction mixture is stirred at room temperature for an additional 3.5 hours, then terminated by dialysis against 2 liters of aqueous triethylamine pH 11. Chromatography of the product on a Sephadex G-75 column (5×100 cm) equilibrated with aqueous triethylamine, pH 11 is then conducted to isolate thyroxine (T4) conjugated to soybean trypsin inhibitor from free T4. Fractions containing 1-2 molecules of thyroxine per molecule of protein are isolated as the preferred reagent. Higher substituted fractions were obtained, although operable, they were less preferable.

1. additionhas been added
2. customterminated by dialysis against 2 liters of aqueous triethylamine pH 11