PTB binding with JEV RNA <i>in vitro</i>. BhullarDeepika JalodiaRicha KaliaManjula VratiSudhanshu 2014 <p>(a) Binding reactions were carried out <i>in vitro</i> using 1 ng radiolabelled JEV RNA (indicated at the bottom) and increasing amounts of recombinant PTB (amounts in ng shown at the top). The RNA-protein complexes were resolved on a 6% non-denaturing PAGE followed by autoradiography. (b) Filter binding assays were carried out by incubating increasing amounts of PTB with 100 fmol <sup>32</sup>P-labelled RNA in binding buffer containing 500 ng yeast tRNA. The percentage of bound RNA was determined and saturation binding curve plotted to calculate the dissociation constant (<i>K</i><sub>d</sub>) of the binding reaction.</p>