Preparation of proteins and peptides for mass spectrometry analysis in a bottom-up proteomics workflow. Curr Protoc Mol Biol 2009 Oct;Chapter 10:Unit10.25
Date
10/10/2009Pubmed ID
19816929Pubmed Central ID
PMC2905857DOI
10.1002/0471142727.mb1025s88Scopus ID
2-s2.0-78651366142 (requires institutional sign-in at Scopus site) 224 CitationsAbstract
This unit outlines the steps required to prepare a sample for MS analysis following protein separation or enrichment by gel electrophoresis, liquid chromatography, and affinity capture within the context of a bottom-up proteomics workflow in which the protein is first broken up into peptides, either by chemical or enzymatic digestion, prior to MS analysis. Also included are protocols for enrichment at the peptide level, including phosphopeptide enrichment and reversed-phase chromatography for sample purification immediately prior to MS analysis. Finally, there is a discussion regarding the types of MS technologies commonly used to analyze proteomics samples, as well as important parameters that should be considered when analyzing the MS data to ensure stringent and robust protein identifications and characterization.
Author List
Gundry RL, White MY, Murray CI, Kane LA, Fu Q, Stanley BA, Van Eyk JEMESH terms used to index this publication - Major topics in bold
Mass SpectrometryPeptides
Proteins
Proteomics
Specimen Handling
