Loxosceles rufescens is the most widely distributed species of the Sicariidae. Its native range spans the semiarid subtropics of the western Palearctic region, as east as Afghanistan. It has also been introduced as a synanthropic species to many localities around the world, including countries in central Europe, East Asia, sub-Saharan Africa, North America as well as Australia, Hawaii and Macaronesia. In Israel it is ubiquitous inside and outside caves in the northern, mesic half of the country. It is found in all cave types, but is more common in dry caves. L. rufescens also found in urban and agricultural settings throughout Israel. 

Loxosceles rufescens exhibits thigmotaxis, and is mostly found in cracks and under stones. It lines the floor of its retreat with silk, but does not construct a web. 

Loxosceles rufescens is considered a medically significant species, as its venom is known to cause necrotic lesions in humans.