Preparation of proteins and peptides for mass spectrometry analysis in a bottom-up proteomics workflow.
Curr Protoc Mol Biol
; Chapter 10: Unit10.25, 2009 Oct.
Article
em En
| MEDLINE
| ID: mdl-19816929
ABSTRACT
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
Manejo de Espécimes
/
Espectrometria de Massas
/
Proteínas
/
Proteômica
Idioma:
En
Revista:
Curr Protoc Mol Biol
Ano de publicação:
2009
Tipo de documento:
Article
País de afiliação:
Estados Unidos