Comparison of Chaotropic Reagents in Peptide Mapping Workflows (TN-1268)
Peptide mapping is a common method for protein characterization. The general workflow includes the isolation of a protein, followed by in solution digest using a serine protease to yield peptides, which are subsequently analyzed by LC and/or MS techniques. Because of its specificity and the general size of peptides generated, trypsin is most commonly used . To optimize sequence coverage, the protein must be denatured prior to trypsin digestion. In this application note, we investigate the differences in results for sequence coverage and overall chromatographic performance using two commonly used chaotropic agents for sample denaturation urea and guanidine HCl.