反应详情

Morphine-HCTL (0.068 g, 1.3×10-4 mol, example 9) was dissolved in 5.7 ml of 70% dimethylformamide/30% water and 1.43 ml 1N potassium hydroxide was added. After 5 min the thiol concentration was determined by reaction with DTNB to be 16.9 mM. The morphine thiol -- (6.4 ml, 1.1×10-4 mol) was added with stirring to 26.7 ml SMCC-BSA, 15/1, (0.16 g, 2.4×10-6 mol). The container was purged with argon and the solution was stirred at room temperature for 2 hr. The protein solution was then dialyzed using a PYROSART module against 10 mM (2-(N-morpholino) ethane sulfonic acid, pH 5.77, and 29.5 ml of 5.39 mg/ml BSA-morphine was collected. The protein solution was frozen at -70° C.

1. concentrationAfter 5 min the thiol concentration
2. customwas determined by reaction with DTNB
3. customThe container was purged with argon
4. customThe protein solution was then dialyzed
5. customwas collected
6. temperatureThe protein solution was frozen at -70° C.