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. 2016 Aug 4;6:31041. doi: 10.1038/srep31041

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Copyright © 2016, The Author(s)

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(a) The concentration of protein used during immobilization to the quartz microbalance and the concentration of soluble protein added during subsequent measurements were both set to 100 ng/μl. The “Baseline” (red) denotes signal changes detected when buffer containing no protein is added to GroEL-AD immobilized sensors. The “α-Syn” (black) and “Aβ42” (blue) signals were measured by adding soluble aliquots of α-Synuclein or Aβ42, respectively, to the reaction chamber containing immobilized GroEL-AD. The “GroES” signal (green), however, was measured by adding soluble GroEL-AD to a reaction chamber containing immobilized GroES protein, in the presence of 0.4 M Gdn-HCl. See the Materials and Methods section for more details. (b) Sensorgrams measured using a quartz microbalance with immobilized GroES and varying concentrations of soluble GroEL-AD in the presence of 0.4 M Gdn-HCl. The concentration of GroEL-AD during each experiment was as follows (from top to bottom); 0 μM, 0.495 μM, 1.23 μM, 2.48 μM, 2.97 μM, 3.71 μM, 4.95 μM, 9.90 μM. Each trace was analyzed using the analysis function of Aqua 2.0 to obtain k_obs and ΔF values. (c) Plot of the estimated |ΔF| values to the concentration of soluble GroEL-AD added. Data points were fitted non-linearly to the isothermal adsorption equation outlined in Materials and Methods to obtain the fitted curve shown in the figure. (d) Linear regression plots of k_obs to the concentration of soluble [GroEL-AD]. We used only the k_obs values for the lower [GroEL-AD] concentrations in this analysis since the data sampling rate, which was fixed for the instrument, precluded the detailed sampling of raw sensorgrams with large k_obs values. This leads to more errors to be incorporated into the k_obs estimates at higher [GroEL-AD] concentrations, and subsequently a notable tendency in the linear regression analysis to yield negative values of k_off (the y-intercept).