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Tag-bound models were generated for each condition, solvated, and energy minimized as above, except the temperature was gradually raised from 300 to 350 K during equilibration and then simulated for 100 ns at 350 K

When the TSA method was used, the sections were treated with biotinylated goat anti-mouse/rabbit IgG (1:1000 dilution, Vector Laboratories, Burlingame, CA) for 1 h, and then incubated with HRP-conjugated-avidin for 30 min (1:100 dilution in TN buffer, TSA System) at room temperature

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