Hello everyone. I want to do trypsin digestion for peptides sample preparation in LC/MS. But my laboratory doesn't have DDT (Dithiothreitol). Can I use b-mercaptoetanol in stead of DDT for reducing disulfide bond of protein before using Trypsin in digestion? And I don't have IAA for alkalinization, too. So what should I do now?
If you have any protocol about using b-mercaptoetanol for peptides sample preparation in LC/MS, please send me that. Thank you!
BME works fine at similar concentrations to DDT. I don't use it anymore because BME gives me a migrane. You actually don't need to cap the resulting C's. As long as you keep the sample reduced (i.e., away from O2) you can use it directly in MS. I used to add a tiny amount of sulfite to the sample before ESI to preserve the reducing environment. However, as long as you can smell the BME you should be fine.
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