Ethanol → N-Hydroxysuccinimide

Participants

Reaction Steps

Rabbit IgG (10 mg) was dissolved in 2 mL of 0.1M sodium phosphate buffer containing 0.1M NaCl (pH 7.5) and treated with an ethanol solution of N-succinimidyl-3-(2-pyridyl-dithio)propionate (SPDP) (2-5 equiv. of 9.6 mM solution). The SPDP solution was added in aliquots over a period of 30 minutes. The unreacted SPDP and N-hydroxy-succinimide formed were removed by gel filtration on Sephadex G-25 column (1×14 cm) that was equilibrated with 0.1M sodium phosphate buffer/0.1M NaCl (pH 7.5) and the SPDP-coupled protein eluted with same buffer.

Provenance & Evidence