Skip to main content
. 2020 Jan 28;48(8):4013–4027. doi: 10.1093/nar/gkaa034

Figure 5.

Figure 5.

Direct ELONA experiments to identify the Sequence 7-bound target protein among a set of recombinant proteins. (A) Absorbance levels of horseradish peroxidase (HRP) activity using 10 nM of the indicated recombinant proteins detected by 20 nM of biotinylated Sequence 7 or Sequence 7RC premixed with streptavidin-poly-HRP. Error bars: S.D.; n = 3 for all proteins except for YBX1 (n = 6). (B) Titration of 10 nM YBX1 with increasing concentrations of Sequence 7 (blue diamonds) versus Sequence 7RC (red squares), and no ssODN (green triangles). The no protein/Sequence 7 (purple cross) and no protein/Sequence 7RC (blue star) controls did not change with concentration. (C) The same experiment carried out using 10 nM RBM3 instead of YBX1. Assuming the curve to be fully saturated at 3.0 absorbance with half-maximal 1.5 absorbance, a KD of approximately 3.0 nM can be estimated. (D) ELONA performed for each of the nine sequences and their respective scrambled version 1 on 10 nM YBX1. Sequences 2 and 7 were also tested with their respective scrambled versions 2. Error bars: S.D. n = 6 for selected sequences 2 and 7, n = 3 for all other sequences.