Phosphate → HCID 53629836, compound

Participants

Reaction Steps

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.).

Provenance & Evidence