반응 #1747849
ord-34cbc220044e4ddfa08a0e8d4e75a805
반응 방정식
반응 조건
후처리
- 1기타to react for at least 24 hours at room temperature
- 2기타To separate fluorescently labeled desferrioxamine from free fluorescein and free desferrioxamine
- 3기타the mixture is separated
- 4세척are eluted in 2 mL fractions
- 5세척The fluorescein-desferrioxamine is eluted in the void volume of the column
- 6기타The first peak is collected
- 7기타used for preparing fluorescein-desferrioxamine, desferrioxamine, and rhodamine liposomes
실험 절차
An aliquot of 120 mg of desferrioxamine and 233.64 mg of fluorescein isothiocyanate (FITC) are mixed in 50 mL of 0.1 M sodium carbonate pH 9.5. A molar ratio of 3:1 of FITC: desferrioxamine is used as described in Hentz et al. (1997) Anal. Chem. 69:4994-5000. The reaction mixture is wrapped in aluminum foil to avoid light exposure and allowed to react for at least 24 hours at room temperature. To separate fluorescently labeled desferrioxamine from free fluorescein and free desferrioxamine, the mixture is separated using a P-2 column (Bio-Rad) equilibrated in saline and samples are eluted in 2 mL fractions. The fluorescein-desferrioxamine is eluted in the void volume of the column. The first peak is collected and used for preparing fluorescein-desferrioxamine, desferrioxamine, and rhodamine liposomes.