Proteolytic digestion in histidine buffer is comparable to data obtained from digestion in ammonium bicarbonate in terms of number of peptides identified and amino acid sequence coverage. More complete separation is observed for tryptic peptides prepared in histidine buffer and separated using the MSWIFT. We attribute the more efficient separation in the MSWIFT due to the low conductivity of histidine and therefore higher field strength across separation wells resulting in the migration of peptides to the proper well in a rapid manner. Sample throughput is increased since shorter separation times are needed. Peptide pI can be used as a validation criteria for peptide identification/assignment since more efficient separations are observed when using histidine buffer.
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