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- aggregation classification "A1".
- aggregation creator person.
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- aggregation date "2011".
- aggregation format "application/pdf".
- aggregation hasFormat 1907040.bibtex.
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- aggregation isPartOf urn:issn:1754-2189.
- aggregation language "eng".
- aggregation rights "I have transferred the copyright for this publication to the publisher".
- aggregation subject "Biology and Life Sciences".
- aggregation title "Selecting protein N-terminal peptides by combined fractional diagonal chromatography".
- aggregation abstract "In recent years, procedures for selecting the N-terminal peptides of proteins with analysis by mass spectrometry have been established to characterize protease-mediated cleavage and protein alpha-N-acetylation on a proteomic level. As a pioneering technology, N-terminal combined fractional diagonal chromatography (COFRADIC) has been used in numerous studies in which these protein modifications were investigated. Derivatization of primary amines-which can include stable isotope labeling-occurs before trypsin digestion so that cleavage occurs after arginine residues. Strong cation exchange (SCX) chromatography results in the removal of most of the internal peptides. Diagonal, reversed-phase peptide chromatography, in which the two runs are separated by reaction with 2,4,6-trinitrobenzenesulfonic acid, results in the removal of the C-terminal peptides and remaining internal peptides and the fractionation of the sample. We describe here the fully matured N-terminal COFRADIC protocol as it is currently routinely used, including the most substantial improvements (including treatment with glutamine cyclotransferase and pyroglutamyl aminopeptidase to remove pyroglutamate before SCX, and a sample pooling scheme to reduce the overall number of liquid chromatography-tandem mass spectrometry analyses) that were made since its original publication. Completion of the N-terminal COFRADIC procedure takes similar to 5 d.".
- aggregation authorList BK854878.
- aggregation endPage "1141".
- aggregation issue "8".
- aggregation startPage "1130".
- aggregation volume "6".
- aggregation aggregates 3147656.
- aggregation isDescribedBy 1907040.
- aggregation similarTo nprot.2011.355.
- aggregation similarTo LU-1907040.