Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift

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dc.contributor.authorLewis, Karen A.
dc.contributor.authorAltschuler, Sarah E.
dc.contributor.authorWuttke, Deborah
dc.date.accessioned2020-03-13T18:10:48Z
dc.date.available2020-03-13T18:10:48Z
dc.date.issued2019-01
dc.description.abstractMeasuring protein/DNA interactions that have apparent binding affinity constants in the low-picomolar range presents a unique experimental challenge. To probe the sequence specificity of telomere binding proteins, our laboratory has developed an electrophoretic mobility shift assay protocol that allows for the routine measurement of K D,app values in the 1-20 pM range. Here, we describe the protocol and highlight the particular considerations that should be made to successfully and reproducibly measure high-affinity interactions between proteins and single-stranded DNA.
dc.description.departmentChemistry and Biochemistry
dc.description.versionThis is the accepted manuscript version of an article published in Methods in Molecular Biology.
dc.formatText
dc.format.extent12 pages
dc.format.medium1 file (.pdf)
dc.identifier.citationLewis, K. A., Altschuler, S. E., & Wuttke, D. S. (2019). Measuring low-picomolar apparent binding affinities by minigel electrophoretic mobility shift. Methods in Molecular Biology, 1855.
dc.identifier.doihttps://doi.org/10.1007/978-1-4939-8793-1_29
dc.identifier.issn1940-6029
dc.identifier.urihttps://hdl.handle.net/10877/9447
dc.language.isoen
dc.publisherHumana Press
dc.sourceMethods in Molecular Biology, 2019, Vol. 1855.
dc.subjectEMSA
dc.subjectDNA binding protein
dc.subjectbinding assay
dc.subjecthigh-affinity binding
dc.subjectnative gel
dc.subjectChemistry and Biochemistry
dc.titleMeasuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift
dc.typeArticle

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