Jumping spiders are known for complex courtship displays with both visual and vibratory components, but increasing evidence shows they also use chemoreception in intraspecific communication. We conducted two experiments using Habronattus pyrrithrix (Chamberlin, 1924) to assess male response to substrate-borne or airborne chemical cues produced by virgin females. First, we tested the effect of substrate-borne cues by allowing males to inspect two pieces of filter paper that had either been exposed to a female (thus covered in silk and/or excreta) or not (control). Second, we used a Y-tube olfactometer to test male response to female airborne cues versus a no-odor control in the absence of substrate-borne cues. Males responded to substrate-borne cues (spending more time traversing and palpating female-treated filter paper compared with the control) but did not respond to airborne cues alone. Together, these experiments suggest male H. pyrrithrix may use contact chemical cues from female silk to locate or assess females.