Immunoprecipitation of mRNA-protein complexes.
Nat Protoc
; 1(2): 577-80, 2006.
Article
em En
| MEDLINE
| ID: mdl-17406284
ABSTRACT
Immunoprecipitation of mRNA-protein complexes is a method that can be used to study RNA binding protein (RBP)-RNA interactions. In this protocol, an antibody targeting an RBP of interest is used to immunoprecipitate the RBP and any interacting molecules from a cell lysate. Reverse transcription followed by PCR is then used to identify individual mRNAs isolated with the RBP. This method focuses on examining an association between a specific RBP-mRNA complex, and it is best suited for a small scale screening of known or putative binding partners. It can also be used as a second, independent method to verify RBP-mRNA interactions discovered through more universal screening techniques. We describe the immunoprecipitation protocol in practical detail and discuss variations of the method as well as issues associated with it. The procedure takes three days to complete.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
RNA Mensageiro
/
Proteínas de Ligação a RNA
/
Imunoprecipitação
Tipo de estudo:
Guideline
Limite:
Animals
Idioma:
En
Revista:
Nat Protoc
Ano de publicação:
2006
Tipo de documento:
Article