| Electrophoretic mobility shift assay (EMSA) for detecting protein-nucleic acid interactions. | |
| | |
MedLine Citation:
|
PMID: 17703195 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The gel electrophoresis mobility shift assay (EMSA) is used to detect protein complexes with nucleic acids. It is the core technology underlying a wide range of qualitative and quantitative analyses for the characterization of interacting systems. In the classical assay, solutions of protein and nucleic acid are combined and the resulting mixtures are subjected to electrophoresis under native conditions through polyacrylamide or agarose gel. After electrophoresis, the distribution of species containing nucleic acid is determined, usually by autoradiography of 32P-labeled nucleic acid. In general, protein-nucleic acid complexes migrate more slowly than the corresponding free nucleic acid. In this protocol, we identify the most important factors that determine the stabilities and electrophoretic mobilities of complexes under assay conditions. A representative protocol is provided and commonly used variants are discussed. Expected outcomes are briefly described. References to extensions of the method and a troubleshooting guide are provided. |
| | |
Authors:
|
Lance M Hellman; Michael G Fried |
Related Documents
:
|
807245 - Products obtained after in vitro reaction of 7,12-dimethylbenz[alpha]anthracene 5,6-oxi... 11097825 - Determination of internucleotide (h)j(hn) couplings by the modified 2d j(nn)-correlated... 19669535 - Electron correlated ab initio study of amino group flexibility for improvement of molec... 6209715 - Condensation of nucleic acids by intercalating aromatic cations. 6661395 - Attachment of rattlesnake venom myotoxin a to sarcoplasmic reticulum: peroxidase conjug... 759605 - The selection and evaluation of new chelating agents for the treatment of iron overload. |
Publication Detail:
|
Type: Journal Article; Research Support, N.I.H., Extramural |
Journal Detail:
|
Title: Nature protocols Volume: 2 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2007 |
Date Detail:
|
Created Date: 2007-08-17 Completed Date: 2007-12-06 Revised Date: 2011-09-26 |
Medline Journal Info:
|
Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
|
Languages: eng Pagination: 1849-61 Citation Subset: IM |
Affiliation:
|
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky 40536-0509, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Bacterial Proteins
/
metabolism Cyclic AMP Receptor Protein / metabolism DNA-Binding Proteins / analysis*, metabolism Electrophoretic Mobility Shift Assay / methods* Escherichia coli / genetics, metabolism Escherichia coli Proteins / metabolism Humans Lac Operon / genetics Lac Repressors O(6)-Methylguanine-DNA Methyltransferase / metabolism Promoter Regions, Genetic RNA-Binding Proteins / analysis* Repressor Proteins / metabolism Transcription Factors / metabolism |
| Grant Support | |
ID/Acronym/Agency:
|
GM-070662/GM/NIGMS NIH HHS; R01 GM070662-01/GM/NIGMS NIH HHS; R01 GM070662-02/GM/NIGMS NIH HHS; R01 GM070662-03/GM/NIGMS NIH HHS; R01 GM070662-04/GM/NIGMS NIH HHS; R01 GM070662-05/GM/NIGMS NIH HHS; R01 GM070662-06/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Bacterial Proteins; 0/Cyclic AMP Receptor Protein; 0/DNA-Binding Proteins; 0/Escherichia coli Proteins; 0/Lac Repressors; 0/LacI protein, E coli; 0/RNA-Binding Proteins; 0/Repressor Proteins; 0/Transcription Factors; 0/crp protein, E coli; EC 2.1.1.63/O(6)-Methylguanine-DNA Methyltransferase |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Two-dimensional gel electrophoresis for identifying proteins that bind DNA or RNA.
Next Document: A fluorescence-based double retrograde tracer strategy for charting central neuronal connections.