Kinesin and microtubules (MTs) are major components of cytoskeleton in cells of living organisms. Kinesin and microtubules together play crucial roles in a wide range of cellular functions, most significantly intracellular transport. Recent developments in bioengineering and biotechnology allows for using these natural molecules as components of molecular machines and molecular robots. In vitro gliding assay has been the best platform to evaluate the potential of these biomolecules for molecular machines.