Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift. Journal Article uri icon

Overview

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.

publication date

  • January 1, 2019

Full Author List

  • Lewis KA; Altschuler SE; Wuttke DS

Other Profiles

Additional Document Info

start page

  • 341

end page

  • 354

volume

  • 1855