反应详情

Amphetamine-ATP (0.095 g, 3×10−4 mol, Example 18) was dissolved in 50 ml of 0.12 M potassium carbonate in 80% methanol/20% water. After 5 min at room temperature the thiol concentration was determined by reaction with DTNB to be 25 mM. The amphetamine thiol (7.6 ml, 1.9×10−4 mol) was added with stirring to 20.3 ml (0.4 g, 5.9×10−6 mol) of SMCC-BSA, 10/1. The pH of the solution was adjusted to 7 with 1 N hydrochloric acid. The container was purged with argon and the solution was stirred at room temperature for 2 hr. The conjugate solution was then dialyzed with a PYROSTART ultrafiltration module against 10 mM morpholineethanesulfonic acid, pH 5.77 and 110 ml of 3.57 mg/ml BSA-amphetamine was collected. The protein solution was frozen at −70° C.

1. concentrationAfter 5 min at room temperature the thiol concentration
2. customwas determined by reaction with DTNB
3. customThe container was purged with argon
4. filtrationThe conjugate solution was then dialyzed with a PYROSTART ultrafiltration module against 10 mM morpholineethanesulfonic acid, pH 5.77 and 110 ml of 3.57 mg/ml BSA-amphetamine
5. customwas collected
6. temperatureThe protein solution was frozen at −70° C.