Biology Reference
In-Depth Information
Fig.
1
Summarized workflow for a label-free approach. Initial sample is obtained and fractionated by GeLC
(SDS-PAGE represented by the white/gray sections, being
x
and
y
hypothetical samples) or MudPIT (peptides
eluted from each salt step on the SCX chromatography are loaded in an RP column prior to injection in the
mass spectrometer). In the data-dependent acquisition approach the most intense peptide ions are selected
and fragmented and acquired MS/MS spectra are searched against a user-defined database
Elution from SCX column is performed in several steps, increasing
the salt concentration. Peptides are then eluted from each step and
loaded in an
in line
reverse-phase (RP) column prior to injection in
the mass spectrometer (Fig.
1
). Another approach largely used in
