Chapter title |
Gel Mobility Shift Assays to Detect Protein-RNA Interactions.
|
---|---|
Chapter number | 12 |
Book title |
Bacterial Regulatory RNA
|
Published in |
Methods in molecular biology, January 2012
|
DOI | 10.1007/978-1-61779-949-5_12 |
Pubmed ID | |
Book ISBNs |
978-1-61779-948-8, 978-1-61779-949-5
|
Authors |
Alexander V. Yakhnin, Helen Yakhnin, Paul Babitzke |
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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