Reaction of NO quenching
Preparation of Fe(II)-DTCS Complex1. Dissolve 278 mg FeSO4, 7H2O (ferrous sulfate heptahydrate) with 20 ml watera) to prepare 50 mM FeSO4 solution.b)2. Dissolve 123 mg DTCS Na with 10 ml watera) to prepare 50 mM DTCS solution.3. Mix 1 ml DTCS Na solution with 8.8 ml buffer solutiona) (pH 7 or higher). Add 200 μl FeSO4 solution just prior to use.c)a) Purge any dissolved oxygen in the water or the buffer by nitrogen gas bubbling for at least 30 minutes prior to dissolving FeSO4.b) The FeSO4 solution can be stored at -20ºC for at least 2 months.c) Fe(II)-DTCS complex is colorless. If the solution is brown, Fe(III)-DTCS may have formed because of dissolved oxygen in the solution. However, the brown solution can still be used for NO trapping.Preparation of NO-Fe(II)-DTCS Complex1. Under argon gas flow, add 200 μl of FeSO4 solution to 9.4 ml buffer solution (pH 7 or higher). Then introduce NO into the solution by bubbling NO gas through a glass capillary for 15 minutes.2. Add 400 μl of DTCS Na solution to the FeSO4 solution, and continue to introduce NO by bubbling for another 5 minutes.3. Remove excess NO with argon gas bubbling for 5 minutes, and store at -20ºC. The NO-Fe(II)-DTCS solution can be stored at -20ºC for at least 2 months in oxygen-free conditions.
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