Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift
dc.contributor.author | Lewis, Karen A. | |
dc.contributor.author | Altschuler, Sarah E. | |
dc.contributor.author | Wuttke, Deborah | |
dc.date.accessioned | 2020-03-13T18:10:48Z | |
dc.date.available | 2020-03-13T18:10:48Z | |
dc.date.issued | 2019-01 | |
dc.description.abstract | Measuring 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.department | Chemistry and Biochemistry | |
dc.description.version | This is the accepted manuscript version of an article published in Methods in Molecular Biology. | |
dc.format | Text | |
dc.format.extent | 12 pages | |
dc.format.medium | 1 file (.pdf) | |
dc.identifier.citation | Lewis, 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.doi | https://doi.org/10.1007/978-1-4939-8793-1_29 | |
dc.identifier.issn | 1940-6029 | |
dc.identifier.uri | https://hdl.handle.net/10877/9447 | |
dc.language.iso | en | |
dc.publisher | Humana Press | |
dc.source | Methods in Molecular Biology, 2019, Vol. 1855. | |
dc.subject | EMSA | |
dc.subject | DNA binding protein | |
dc.subject | binding assay | |
dc.subject | high-affinity binding | |
dc.subject | native gel | |
dc.subject | Chemistry and Biochemistry | |
dc.title | Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift | |
dc.type | Article |