Phosphate → HCID 53629836, compound

HRID 2374658Notes

Reaction equation

HRID 2374658 reaction equation
P([O-])([O-])([O-])=O.[Na+].[Cl-:7].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O.Cl.[NH2:29][O:30][CH2:31][C:32]([O:34][N:35]1[C:39](=[O:40])[CH2:38][CH2:37][C:36]1=[O:41])=[O:33].CN(C=O)C>CN(C=O)C>[ClH:7].[NH2:29][O:30][CH2:31][C:32]([O:34][N:35]1[C:39](=[O:40])[CH2:38][CH2:37][C:36]1=[O:41])=[O:33]

Participants

6

Reactant 4

HCID 1061PhosphateHCID 5234Sodium ChlorideHCID 6049Ethylenediaminetetraacetic AcidHCID 131724605HCID 131724605, compound

Product 1

HCID 53629836HCID 53629836, compound

Reagents, catalysts & solvents 1

HCID 6228DimethylformamideSolvent

Procedure

A 5 mg/mL solution of ovalbumin in PBS (100 mM phosphate, 150 mM NaCl, pH 7.4) and 2 mM EDTA (200 μL; 1 mg protein) is prepared. A solution of succinimidyl aminooxyacetate hydrochloride (SAAH)(2 mg) in DMF (50 μL) is prepared. To the protein solution is added the SAAH/DMF solution (15 equivalents). The reaction mixture is incubated at room temperature for 4 hours. The modified protein is isolated and buffer exchanged by placing the reaction mixture in a 30K ultra-free centrifugation device and washing three times with 0.1 M MES, 0.90% NaCl, pH 4.7 (3×400 μL). The purified protein is quantified for protein concentration (BCA assay; Pierce Chemical Co., Rockford, Ill.).

Workup

  1. customThe modified protein is isolated
  2. washwashing three times with 0.1 M MES, 0.90% NaCl, pH 4.7 (3×400 μL)
  3. concentrationThe purified protein is quantified for protein concentration (BCA assay; Pierce Chemical Co., Rockford, Ill.)

Sources & evidence

ORD record
ord-ae8a165e71f24639abc11ab9d5c1f24a
Source dataset
uspto-grants-2008_12
Patent number
US07462689B2

Reaction notes

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