I intend to use maleimide chemistry to attach a short peptide with a terminal cysteine to the N-terminal domain of another protein. Do I need to pretreat the short peptide with a reducing agent to eliminate inter-peptide disulfide bonds, or will this not be favored?
Thanks!
Did you synthesize the peptide or did you buy it?, what is the pH of the solution? If you bought the peptide it should already be reduced.
Thanks Gary, I don't have the peptide yet but I intend to order it. I haven't ordered peptide before, but assuming they send it to me lyophilized, do I need to worry about them forming disulfide bonds if reconstituted in neutral PBS? I suppose I could add EDTA to prevent it happening if they are already reduced.
Disulfide bond formation is a possibility. You can keep the Cys residues reduced using TCEP. This will not interfere with the reaction with maleimides.
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