Lewis, Karen A.Altschuler, Sarah E.Wuttke, Deborah2020-03-132020-03-132019-01Lewis, K. A., Altschuler, S. E., & Wuttke, D. S. (2019). Measuring low-picomolar apparent binding affinities by minigel electrophoretic mobility shift. Methods in Molecular Biology, 1855.1940-6029https://hdl.handle.net/10877/9447Measuring 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.Text12 pages1 file (.pdf)enEMSADNA binding proteinbinding assayhigh-affinity bindingnative gelChemistry and BiochemistryArticlehttps://doi.org/10.1007/978-1-4939-8793-1_29