反応 #7437

ord-34edcaa7e3ba4173928e329654c5e7d9

反応方程式

O=C([O-])O
bicarbonate
O=C(CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21)NCCCCCC(=O)ON1C(=O)CC(S(=O)(=O)[O-])C1=O.[Na+]
sulfo-NHS-LC-biotin
→
O=C(O)CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21
Biotin

反応物

O=C([O-])O
bicarbonate
O=C(CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21)NCCCCCC(=O)ON1C(=O)CC(S(=O)(=O)[O-])C1=O.[Na+]
sulfo-NHS-LC-biotin

試薬

なし

溶媒

O
water

反応条件

詳細条件
See reaction.notes.procedure_details.

後処理

  1. 1
    その他to give a final volume of 1.0 mL
  2. 2
    その他to give a 1 mg/mL solution just
  3. 3
    workup.ADDITIONto addition
  4. 4
    workup.ADDITIONThe first aliquot (10 uL) is added
  5. 5
    workup.ADDITIONThe second aliquot (10 uL) is added
  6. 6
    その他The unreacted biotin is removed by size exclusion chromatography

実験手順

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.

出典

DOI: 10.6084/m9.figshare.5104873.v1特許: US07087395B1uspto-grants-2006_08