For the step size and dwell time determinations, Qdot 525 conjugated with goat F(ab’)2 antimouse immunoglobulin G (H + L) were mixed with anti-c-Myc monoclonal antibody (Clontech) as described previously (33 (link), 34 (link)). F-actin was labeled by Alexa Fluor 568 Phalloidin at a 1:1 ratio overnight and stored at 4 °C. For single-molecule experiments with Qdot-labeled HM19, 7.5 μM labeled and unlabeled actins were mixed at the ratio of 1:10, and the mixture was diluted to 1 μM just before the experiments.
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