This medium-sized rattlesnake ranges primarily east of the Rocky Mountains from Canada down to Mexico. In a few states and Canadian provinces, it is the only venomous snake species. It has one of the largest geographic ranges of any rattlesnake species subspecies, behind Crotalus horridus.

The prairie rattlesnake primarily feeds on mammals as adults, but given their diverse habitat and range, this rattlesnake has also been recorded consuming birds, lizards, and amphibians. They can live 16-20 years in the wild, however, only a limited number of young will make it to full adulthood. Their primary predators are birds, such as hawks and roadrunners, while bobcats and skunks are known to occasionally prey on prairie rattlesnakes.

The venom of the prairie rattlesnake primarily consists of myotoxins which leads to muscle necrosis and paralysis. This species is known to hybridize with the Mojave Rattlesnake (Crotalus scutalatus) in the Peloncillo Mountains of New Mexico, which is likely the cause of prairie rattlesnakes in that area possessing a neurotoxic venom.

Juvenile prairie rattlesnakes have been documented to control their venom from birth, contradicting the myth that baby rattlesnakes cannot control their venom output. Juvenile rattlesnakes have to learn from experience how much venom to input into smaller or larger prey, however.