Ticks display a distinct type of host-seeking behaviour called questing. It has been proposed that the questing behaviour of Ixodes scapularis explains the geographic variation in Lyme disease (LD) risk in the eastern USA because the northern population has been shown to quest more often than the southern population. The height at which questing occurs is variable and this study aimed to characterize questing height for I. scapularis. Ticks were collected from a northern and southern state (i.e. Maryland and Texas) and bioassays were conducted. We report that nymphs from Texas quested at lower heights compared to nymphs from Maryland. In addition, only Texas nymphs exhibited a behaviour we call ‘hiding behaviour’. These results may reflect the different composition of hosts between these two areas as the south has a higher abundance of lizards. In contrast, there was no significant difference in questing height between Maryland adults and Texas adults which was to be expected since adults are feeding on white-tailed deer in both locations. If all southern I. scapularis nymphs are questing at lower heights, this might make them less likely to come into contact with humans and this may be contributing to the geographical difference in LD prevalence.