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Peptide fractionation by acid pH SDS‐free electrophoresis
Author(s) -
Ramos Yassel,
Garcia Yairet,
PérezRiverol Yasset,
Leyva Alejandro,
Padrón Gabriel,
Sánchez Aniel,
CastellanosSerra Lila,
González Luis J.,
Besada Vladimir
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201000677
Subject(s) - chromatography , proteome , chemistry , peptide , fractionation , electrophoresis , histidine , polyacrylamide gel electrophoresis , free flow electrophoresis , gel electrophoresis , gel electrophoresis of proteins , amino acid , biochemistry , enzyme
SDS‐free polyacrylamide gel electrophoresis is an effective alternative approach to peptide fractionation. Here we describe a discontinuous buffer system at acid pH that improves the separation of acidic peptides from tryptic digestion. MOPS and chloride act as trailing and leading ions, respectively, in this system, while histidine operates as counterion and buffers all solutions. In these electrophoretic conditions, peptides with p I below 5.5 migrate with low overall electrophoretic mobilities but high differences from one another, which allows for their efficient resolution . In silico analysis of several proteomes shows that the acid pH system allows a peptide simplification of 2.5‐fold with respect to the total peptide mixture, and still a proteome coverage of about 95% is achievable. A straightforward method with a protocol including proteomic studies was achieved for SDS‐PAGE of proteins, enzyme treatment and further peptide fractionation by SDS‐free acid PAGE.