The PDF file includes:

• Fig. S1. Binding transitions obtained by HS-AFM.
• Fig. S2. Dwell time distributions and the fitted time course of P_bound.
• Fig. S3. Simulated binding transitions.
• Fig. S4. The AgTx2 concentration dependence on k_obs.
• Fig. S5. Boltzmann’s H function.
• Fig. S6. Dynamic disorder of binding transitions and correlation of individual P_bound values and interchannel distance.
• Fig. S7. Dwell time distributions obtained by dwell time analysis, event-oriented analysis, and correlation function.
• Fig. S8. Removing fluctuations in the baseline.
• Reference (61)

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Other Supplementary Material for this manuscript includes the following:

• Movie S1 (.mp4 format). HS-AFM movie of AgTx2 binding on the KcsA channels.
• Movie S2 (.mp4 format). HS-AFM movie showing dynamic disorder of binding transitions.