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Concentrate the antibody to 5-7 mg/ml
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Dissolve 10 mg of SATA in 1 ml DMF (SATA:DMF)
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Add 3.0 m l SATA:DMF per mg of antibody
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Gently stir solution for 2 hours at room temperature.
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Dissolve 0.5 g hydroxylamine hydrochloride in 10 ml PBS:EDTA
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Add 0.25 g NaOH pellets to this solution to neutralize to approximately pH 7.0
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Add to antibody solution at 6.49 m l/m l SATA.
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Gently stir for 30 minutes at room temperature.
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Equilibrate column with PBS:EDTA.
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Desalt thiolated antibody on column. Collect in the smallest volume possible without collecting any SATA. Allow free SATA (2nd peak) to come off the column.
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Store the thiolated antibody at 2-8o C for no more than one hour.
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Re-equilibrate the column with PBS.