In the wolf spider, Hogna helluo, we tested the response to insect and spider prey chemical cues and whether they show a preference for cues associated with prey consumed most recently. Thirty adult female H. helluo were maintained on a diet of either females of a smaller co-occurring wolf spider (Pardosa milvina) or domestic crickets (Acheta domesticus). A single P. milvina or cricket nymph was maintained on filter paper for 24 h, after which the papers from both prey sources were simultaneously presented to individual H. helluo from each diet treatment group. H. helluo locomotor behavior on each treatment and initial substrate preference was recorded (n = 15/treatment). H. helluo fed crickets showed significantly longer residence time and decreased mobility on filter paper previously occupied by a cricket; spiders fed P. milvina showed longer residence times and decreased mobility on filter paper previously occupied by P. milvina. H. helluo fed P. milvina exhibited an initial preference for substrates previously occupied by P. milvina but H. helluo fed crickets did not show a corresponding initial preference for crickets. Results suggest that H. helluo can detect distant cues associated with P. milvina but not crickets before contacting the substrate and that H. helluo respond to chemical cues from prey and show a preference for those cues associated with their most recent prey.