Gel mobility shift assays to detect protein-RNA interactions.
Methods Mol Biol
; 905: 201-11, 2012.
Article
em En
| MEDLINE
| ID: mdl-22736005
ABSTRACT
The gel mobility shift assay is a powerful technique for detecting and quantifying protein-RNA interactions. While other techniques such as filter binding and isothermal titration calorimetry (ITC) are available for quantifying protein-RNA interactions, gel shift analysis provides the added advantage that you can visualize the protein-RNA complexes. In the gel shift assay, protein-RNA complexes are typically separated from the unbound RNA using native polyacrylamide gels in Tris/borate/EDTA buffer, although an alternative Tris-glycine buffering system is superior in many situations. Here, we describe both gel shift methods, along with strategies to improve separation of protein-RNA complexes from free RNA, which can be a particular challenge for small RNA binding proteins.
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Base de dados:
MEDLINE
Assunto principal:
RNA
/
Proteínas de Ligação a RNA
/
Ensaio de Desvio de Mobilidade Eletroforética
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article