Reduction of Disulfides in Protein Molecules Using DTT

Complete Reduction of Disulfides in Protein Molecules Using DTT 

Dissolve a disulfide-containing protein or peptide at a concentration of l-10mg/ml in 6M guanidine hydrochloride, 0.01 M sodium phosphate, 0.15 M NaCl, pH 7.4. Alternative 

Incubate for 2 hours at room temperature. For some buried disulfides to become exposed and fully reduced, it may be necessary to heat the solution (in a capped test tube) at 50°C for 30 minutes. Some procedures use a 2-minute incubation in a boiling water bath to completely denature the protein. 

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