Abstract
Electrophoretic mobility shift assays (EMSA) are used to characterize interactions between nucleic acids and proteins in native conditions. This is based on the fact that the electrophoretic mobility of a nucleic acid becomes slower when it forms complexes with proteins. There are many different variants and applications of this methodology. In this chapter we describe a detailed EMSA protocol applied to the study of asthma.
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References
Isidoro-Garcia M, Sanz C, Garcia-Solaesa V et al (2011) PTGDR gene in asthma: a functional, genetic, and epigenetic study. Allergy 66:1553–1562
Oguma T, Palmer LJ, Birben E et al (2004) Role of prostanoid DP receptor variants in susceptibility to asthma. N Engl J Med 351:1752–1763
Revzin A (1989) Gel electrophoresis assays for DNA-protein interactions. Biotechniques 7:346–355
Scientific T (2012) http://www.piercenet.com/proteomics/browse.cfm?fldid=7edac33e-3981-4e58-8a57-057a8280c68f
Fried M, Crothers DM (1981) Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res 9:6505–6525
Roche Applied Science MA (2006) Instruction manual. DIG gel shift kit, 2nd generation (http://www.protocol-online.org/forums/uploads/monthly_01_2010/post-15212-1263167826.ipb)
Russell SA (2001) Molecular cloning. A laboratory manual, vol 2. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA
Reed KC, Mann DA (1985) Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res 13:7207–7221
Didenko VV, Hornsby PJ (1996) A quantitative luminescence assay for nonradioactive nucleic acid probes. J Histochem Cytochem 44:657–660
Hellman LM, Fried MG (2007) Electrophoretic mobility shift assay (EMSA) for detecting protein-nucleic acid interactions. Nat Protoc 2:1849–1861
Gaudreault M, Gingras ME, Lessard M et al (2009) Electrophoretic mobility shift assays for the analysis of DNA-protein interactions. Methods Mol Biol 543:15–35
Williams TL, Levy DL (2013) Assaying cooperativity of protein-DNA interactions using agarose gel electrophoresis. Methods Mol Biol 1054:253–265
Powell L (2013) Analysis of DNA-protein interactions using PAGE: band-shift assays. Methods Mol Biol 1054:237–251
Acknowledgements
This work was supported by the project “Efecto del Ácido Retinóico en la enfermedad alérgica. Estudio transcripcional y su traslación a la clínica,” PI13/00564, integrated into the “Plan Estatal de I + D + I 2013–2016” and cofunded by the “ISCIII-Subdirección General de Evaluación y Fomento de la investigación” and the European Regional Development Fund (FEDER).
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García-Solaesa, V., Sanz-Lozano, C.S. (2016). Interactions of DNA and Proteins: Electrophoretic Mobility Shift Assay in Asthma. In: Isidoro García, M. (eds) Molecular Genetics of Asthma. Methods in Molecular Biology, vol 1434. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3652-6_7
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DOI: https://doi.org/10.1007/978-1-4939-3652-6_7
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-3652-6
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