Optimized Affinity Capture of Yeast Protein Complexes.
Cold Spring Harb Protoc
; 2016(7)2016 07 01.
Article
em En
| MEDLINE
| ID: mdl-27371596
ABSTRACT
Here, we describe an affinity isolation protocol. It uses cryomilled yeast cell powder for producing cell extracts and antibody-conjugated paramagnetic beads for affinity capture. Guidelines for determining the optimal extraction solvent composition are provided. Captured proteins are eluted in a denaturing solvent (sodium dodecyl sulfate polyacrylamide gel electrophoresis sample buffer) for gel-based proteomic analyses. Although the procedures can be modified to use other sources of cell extract and other forms of affinity media, to date we have consistently obtained the best results with the method presented.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
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Cromatografia de Afinidade
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Proteínas de Saccharomyces cerevisiae
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Substâncias Macromoleculares
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Complexos Multienzimáticos
Idioma:
En
Revista:
Cold Spring Harb Protoc
Ano de publicação:
2016
Tipo de documento:
Article