Habitat loss and fragmentation are among the greatest threats to wildlife and biodiversity. Reptiles are particularly susceptible to these threats due to high site fidelity, large home ranges, and slow movement rates. To understand behavioral responses of Eastern Milksnakes (Lampropeltis triangulum) to fragmentation, we compared home range size and movement rates between a fragmented habitat and an intact habitat. Additionally, we quantified road avoidance and habitat selection in the fragmented habitat. In 2015 and 2016, we collected 453 locations from 17 individuals from Rouge National Urban Park (RNUP), the fragmented study area, using radio-telemetry. We compared our results to a previous study with 1,001 locations from 30 individuals at Queen's University Biological Station (QUBS), our intact study area, collected from 2003 to 2004. We found that home ranges were smaller, but daily movement rate (DMD) and distance-per-move (DPM) were greater in the fragmented study area. We also observed that road crossings by snakes occurred less than expected, suggesting active avoidance of roads. Milksnakes in the fragmented habitat selected locations with a greater number of cover objects within open patches surrounded by high density vegetation, which is consistent with previous findings from the intact habitat. Our findings suggest that Eastern Milksnakes benefit from heterogeneous microhabitats and an abundance of available anthropogenic or natural cover.