Bio-Rad PROTEAN® i12™ IEF System - Manual (Page 54)

PROTEAN i12 IEF System Instruction Manual 50 |
Appendix D
IEF for Peptide Fractionation
Prior to LC-MS
IEF fractionation of proteolytically digested samples may be performed prior to analysis by reversed-phase
liquid chromatography coupled to mass spectrometry with electrospray ionization (LC-MS) in “bottom-up” or
“shotgun” proteomics experiments. IEF and reversed-phase LC are fully orthogonal separation techniques,
and combining them in a two-dimensional peptide separation workflow greatly increases the depth of
proteome coverage compared to reversed-phase LC alone (Cargile et al. 2005). The IEF dimension can
be conveniently run on IPG strips using the PROTEAN
® i12 ™ IEF cell. Details on sample preparation, run
conditions, peptide elution, and preparation for LC-MS may be found in bulletin 6140.