Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift
MetadataShow full metadata
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.
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.