The experiment was conducted to compare the MSE and IM-MSE acquisiton modes as well as to find the optimal on column loading. This was done using 1DLC and cytosolic e coli digest.Raw data were processed and searched using Proteinlynx Global Server version 3.0. The following parameters were used to generate peak lists: minimum intensity for precursors was set to 140, minimum intensity for fragment ions 30 and total bin-count was set to minimum of 400. Processed data was searched against the UniprotKB, Escherichia coli K12 strain, concatenated with 125 common laboratory contaminates. Fixed modification was set to carbamidomethylation of C and variable modification was set to oxidation of M. Searches were conducted against a froward and reverse sequence database. Minimum identification criteria included 3 fragment ions per peptide, 5 fragment ions per protein, minimum peptide identification score of 5.9 and minimum of 2 peptides per protein. Search results and spectra were imported into Scaffold and the global false discovery rate was less than 1%.