反应详情

20 mg cyanogen bromide (CNBr) dissolved in 200 μl deionized water were added to 1 ml of 30 mg/ml chondroitin 4-sulfate (CSA). The pH was adjusted to pH 11 by addition of 5 M NaOH. Activated CSA is applied to a PD-10 column and eluded with 200 mM sodium borate buffer, pH 8.0. CSA fractions were directly dropped into 400 μl of 10 mg/ml fluoresceinamine in 200 mM sodium borate buffer, pH 8.0. The mixture was kept overnight at 4° C. for the coupling reaction. Subsequently, the mixture was concentrated to 1 ml using a SpeedVac concentrator and purified on a PD-10 column and eluded by 50 mM sodium acetate buffer, pH 5.5. Fluoresceinamine-labeled CSA (FL-CSA) fractions were collected and their concentration was determined using the Blyscan glycosaminoglycan assay kit (Biocolor Ltd., Newtown Abbey, Northern Ireland).

1. customThe mixture was kept overnight at 4° C. for the coupling reaction
2. concentrationSubsequently, the mixture was concentrated to 1 ml
3. custompurified on a PD-10 column
4. customwere collected
5. concentrationtheir concentration