Table 1 Protein fractionation, peptide separation and mass spectrometric identification strategies for enhancement of proteome identification coverage explored in this study.
From: Protein abundance profiling of the Escherichia coli cytosol
Steps | Procedures |
|---|---|
(A) Protein fractionation | (1) SDS-PAGE slicing (2) Serial ultrafiltration (3) No protein fractionation |
(B) Tryptic digestion | (1) In-solution digestion (2) In-gel digestion |
(C) Peptide chromatography | (1) Strong cation exchange chromatograhpy (2) Strong anion exchange chromatograhpy (3) C18 ion pair chromatograhpy (4) PSDVB with NH4OH, using StageTip (5) No peptide chromatography |
(D) Parent ion selection in LC-MS | (1) Simple repetition (2) Sequential static exclusion (3) Different ion pair reagents in subsequent runs (4) Subdivided scan range (5) Shallow gradient elution |
(E) CID for MS/MS | (1) Quadrupole-TOF (2) Linear ion trap |