반응 #7437
ord-34edcaa7e3ba4173928e329654c5e7d9
반응 방정식
bicarbonate
sulfo-NHS-LC-biotin
→
Biotin
용매
반응 조건
상세 조건
See reaction.notes.procedure_details.
후처리
- 1기타to give a final volume of 1.0 mL
- 2기타to give a 1 mg/mL solution just
- 3workup.ADDITIONto addition
- 4workup.ADDITIONThe first aliquot (10 uL) is added
- 5workup.ADDITIONThe second aliquot (10 uL) is added
- 6기타The unreacted biotin is removed by size exclusion chromatography
실험 절차
About 2.0 mg of DBP is buffer exchanged with 20 mM bicarbonate at pH 9.6 three times to give a final volume of 1.0 mL. Two aliquots of sulfo-NHS-LC-biotin (Pierce) are dissolved in water to give a 1 mg/mL solution just prior to addition. The first aliquot (10 uL) is added and the reaction is maintained at room temperature for 5 minutes. The second aliquot (10 uL) is added and the reaction proceeds for an additional 4 minutes. The unreacted biotin is removed by size exclusion chromatography using SEPHADEX G-25 and 100 mM PBS, pH 7.4 as the mobile phase.