Biochemical approaches including the design and use of strains expressing epitope-tagged proteins.
Methods Cell Biol
; 109: 347-55, 2012.
Article
em En
| MEDLINE
| ID: mdl-22444151
ABSTRACT
Epitope tagging is a powerful approach used to enable investigations of a cellular component by elucidating its localization, interaction partners, and/or activity targets. Successful tag-based affinity purification yields a mixture of the molecule of interest, associated proteins and nucleic acids, and nonspecific background proteins and nucleic acids, many of which can depend on details of the protocol for enrichment. This chapter provides guidelines and considerations for designing an affinity purification experiment, beginning with construction of a strain expressing a tagged subunit. Common biochemical methods for detecting protein, RNA, and DNA in Tetrahymena thermophila are also discussed.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Marcadores de Afinidade
/
Proteínas de Protozoários
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Técnicas Imunológicas
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Tetrahymena thermophila
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Mapeamento de Epitopos
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Epitopos
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article