Quantification of Spatial Patterns of Microtubule Transport by Kinesin-1 Head and Tail.

kinesin-1 constructs: the microtubule binding tail and isoleucine-alanine-lysine (IAK)-deleted kinesin-1 full length. • This protocol describes a typical gliding assay setup for the kinesin motor domain, alone as well as in combination with the kinesin tail. • We present a systematic framework for typical gliding assays, including experimental acquisition as well as quantitative analysis of microtubule collective transport. • This protocol gives a quantitative metric for microtubule spatial patterns, which enables systematic analysis of microtubule curvature, both in vitro and in vivo.