Protein microarrays for identification of novel extracellular protein-protein interactions.
Curr Protoc Protein Sci
; Chapter 27: Unit 27.3, 2013 Apr.
Article
em En
| MEDLINE
| ID: mdl-23546621
ABSTRACT
Functional protein microarrays offer the capability for high-throughput protein interaction analysis and have long promised to be a powerful tool for understanding protein interactions at the proteome scale. Although popular techniques for protein-protein interaction mapping like yeast-two-hybrid and affinity-purification mass spectrometry have performed well for identifying intracellular protein-protein interactions, the study of interactions between extracellular proteins has remained challenging for these methods. Instead, the use of protein microarrays appears to be a robust and efficient method for the identification of interactions among the members of this class of protein. This unit describes methods for extracellular protein microarray production, screening, and analysis. A protocol is described for enhanced detection of low-affinity interactions by generating multivalent complexes using Fc-fusion bait proteins and protein A microbeads, along with a statistical method for hit scoring and identification of nonspecific interactions.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Mapeamento de Interação de Proteínas
/
Análise Serial de Proteínas
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article