Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Horizontal Gel Electrophoresis for Enhanced Detection of Protein-RNA Complexes.
Dowdle ME
,
Imboden SB
,
Park S
,
Ryder SP
,
Sheets MD
.
???displayArticle.abstract???
Native polyacrylamide gel electrophoresis is a fundamental tool of molecular biology that has been used extensively for the biochemical analysis of RNA-protein interactions. These interactions have been traditionally analyzed with polyacrylamide gels generated between two glass plates and samples electrophoresed vertically. However, polyacrylamide gels cast in trays and electrophoresed horizontally offers several advantages. For example, horizontal gels used to analyze complexes between fluorescent RNA substrates and specific proteins can be imaged multiple times as electrophoresis progresses. This provides the unique opportunity to monitor RNA-protein complexes at several points during the experiment. In addition, horizontal gel electrophoresis makes it possible to analyze many samples in parallel. This can greatly facilitate time course experiments as well as analyzing multiple reactions simultaneously to compare different components and conditions. Here we provide a detailed protocol for generating and using horizontal native gel electrophoresis for analyzing RNA-Protein interactions.
???displayArticle.pubmedLink???
28784977
???displayArticle.pmcLink???PMC5613780 ???displayArticle.link???J Vis Exp ???displayArticle.grants???[+]
Buczek,
Thermodynamic characterization of binding Oxytricha nova single strand telomere DNA with the alpha protein N-terminal domain.
2006, Pubmed
Buczek,
Thermodynamic characterization of binding Oxytricha nova single strand telomere DNA with the alpha protein N-terminal domain.
2006,
Pubmed
Chicoine,
Bicaudal-C recruits CCR4-NOT deadenylase to target mRNAs and regulates oogenesis, cytoskeletal organization, and its own expression.
2007,
Pubmed
Dahlberg,
Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels.
1969,
Pubmed
Foley,
A Platform to Enable the Pharmacological Profiling of Small Molecules in Gel-Based Electrophoretic Mobility Shift Assays.
2016,
Pubmed
Gamberi,
The Bic-C family of developmental translational regulators.
2012,
Pubmed
Hellman,
Electrophoretic mobility shift assay (EMSA) for detecting protein-nucleic acid interactions.
2007,
Pubmed
Maisonneuve,
Bicaudal C, a novel regulator of Dvl signaling abutting RNA-processing bodies, controls cilia orientation and leftward flow.
2009,
Pubmed
,
Xenbase
Pagano,
RNA recognition by the embryonic cell fate determinant and germline totipotency factor MEX-3.
2009,
Pubmed
Pagano,
Quantitative approaches to monitor protein-nucleic acid interactions using fluorescent probes.
2011,
Pubmed
Royer,
Fluorescence approaches to quantifying biomolecular interactions.
2008,
Pubmed
Ryder,
Quantitative analysis of protein-RNA interactions by gel mobility shift.
2008,
Pubmed
Saffman,
Premature translation of oskar in oocytes lacking the RNA-binding protein bicaudal-C.
1998,
Pubmed
Su,
Electrophoresis of proteins and protein-protein complexes in native polyacrylamide gels using a horizontal gel apparatus.
1994,
Pubmed
Tran,
The RNA-binding protein bicaudal C regulates polycystin 2 in the kidney by antagonizing miR-17 activity.
2010,
Pubmed
,
Xenbase
Zhang,
Bicaudal-C spatially controls translation of vertebrate maternal mRNAs.
2013,
Pubmed
,
Xenbase
Zhang,
Determinants of RNA binding and translational repression by the Bicaudal-C regulatory protein.
2014,
Pubmed
,
Xenbase
