biostudies-literatureWebb KJNCRR NIH HHSNHLBI NIH HHSNIMH NIH HHSNIGMS NIH HHS2177-84https://www.ebi.ac.uk/biostudies/studies/S-EPMC3815592biostudies-literatureUnknown12(5)A modified multidimensional protein identification technology (MudPIT) separation was coupled to an LTQ Orbitrap Velos mass spectrometer and used to rapidly identify the near-complete yeast proteome from a whole cell tryptic digest. This modified online two-dimensional liquid chromatography separation consists of 39 strong cation exchange steps followed by a short 18.5 min reversed-phase (RP) gradient. A total of 4269 protein identifications were made from 4189 distinguishable protein families from yeast during log phase growth. The "Micro" MudPIT separation performed as well as a standard MudPIT separation in 40% less gradient time. The majority of the yeast proteome can now be routinely covered in less than a days' time with high reproducibility and sensitivity. The newly devised separatJournal of proteome researchModified MuDPIT separation identified 4488 proteins in a system-wide analysis of quiescence in yeast.PMC3815592R01 MH067880-09P41 RR011823/GM103533HHSN268201000035CP41 RR011823P41 GM103533R01 MH067880Yates JRWebb KJXu TPark SKfalseModified MuDPIT separation identified 4488 proteins in a system-wide analysis of quiescence in yeast.A modified multidimensional protein identification technology (MudPIT) separation was coupled to an LTQ Orbitrap Velos mass spectrometer and used to rapidly identify the near-complete yeast proteome from a whole cell tryptic digest. This modified online two-dimensional liquid chromatography separation consists of 39 strong cation exchange steps followed by a short 18.5 min reversed-phase (RP) gradient. A total of 4269 protein identifications were made from 4189 distinguishable protein families from yeast during log phase growth. The "Micro" MudPIT separation performed as well as a standard MudPIT separation in 40% less gradient time. The majority of the yeast proteome can now be routinely covered in less than a days' time with high reproducibility and sensitivity. The newly devised separat2013-01-01T00:00:00Z2013 May2025-04-18T14:07:54.819Z2019-03-27T01:17:59ZS-EPMC38155922354044610.1021/pr400027m