Реакция #164456

Уравнение реакции

NADH

Bromoacetic acid

Bromoacetic acid

DMF

bromoacetic acid

carbonate

→

Bromoacetyl Glucose-6-Phosphate

Растворители

phosphate carbonate

Условия реакции

Обработка

1. 1
   workup.ADDITIONafter each addition
2. 2
   Другоеwas obtained
3. 3
   ДругоеG6PDH conjugation solution was dialyzed with 3×4 liter portions of 0.01 M phosphate, pH 7.2

Методика

100 μL DMF was added to bromoacetic acid NHS (Sigma 3.06 mg, 12.97 μM) and stirred. A 2.0 mL (10 mg/mL) G6PDH solution was prepared in 0.025 M phosphate carbonate buffer, pH 7.2 and adjusted to pH 8.5 with 0.4 M carbonate buffer. 45 mg disodium G6P and 90 mg NADH, was dissolved in the G6PDH solution. Bromoacetic acid NHS was added to G6PDH solution at 5 μL increments. Enzyme activity was measured on the HITACHI 917 analyzer after each addition. Bromoacetic acid NHS was added until approximately 63.0% enzyme deactivation was obtained. G6PDH conjugation solution was dialyzed with 3×4 liter portions of 0.01 M phosphate, pH 7.2.

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