No peptide left behind: the “out of range” recovery in IPG–IEF fractionation

IEF is often used in multidimensional shotgun proteomics and the narrow range of 3.5–4.5 is the recommended pH interval for the fractionation of tryptic peptides. Usually, even if IEF is performed in IPG strip with a narrow range pH, the entire sample must be loaded onto the strip, including the “ou...

Full description

Saved in:
Bibliographic Details
Published in:Amino acids 2014-05, Vol.46 (5), p.1415-1417
Main Authors: Iori, Elisabetta, Rattazzi, Marcello, Millioni, Renato
Format: Article
Language:English
Subjects:
Analytical Chemistry
Biochemical Engineering
Biochemistry
Biomedical and Life Sciences
fractionation
Hydrogen-Ion Concentration
Isoelectric Focusing - instrumentation
Isoelectric Focusing - methods
Life Sciences
Neurobiology
peptides
Peptides - chemistry
proteins
Proteomics
Proteomics - instrumentation
Proteomics - methods
Short Communication
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:IEF is often used in multidimensional shotgun proteomics and the narrow range of 3.5–4.5 is the recommended pH interval for the fractionation of tryptic peptides. Usually, even if IEF is performed in IPG strip with a narrow range pH, the entire sample must be loaded onto the strip, including the “out of IPG range” peptides. We describe a simple protocol to recover at least a part of these missing peptides and show that this recovery significantly influences the overall fractionation result, increasing the number of the identified proteins and the protein coverage.
ISSN:0939-4451
1438-2199
DOI:10.1007/s00726-014-1720-4