Some cats experience stress when they have novel experiences, such as infrequent transport. This study was a randomized, placebo-controlled, blinded study that sought to objectively evaluate the effects of a 2M2B collar on transported cat physiology and behavior. The statistical model included effects of cat treatment (2M2B vs. control), period (70 min), sex, and interactions. Cats wearing 2M2B collars had an 8% lower PR (p < 0.01), and they slept more and did not hide at the back of the kennel. While control cats vomited or showed excess salivation, cats with 2M2B collars did not show these signs of stress. Male cats were less active during transport than females. Male cats slept more with 2M2B collars compared with male cats with a control collar, but females showed similar sleeping overall regardless of which collar they wore. Female cats increased activity during transport when they had a 2M2B collar, while male activity did not differ with control or 2M2B collars. These data support the concept that the semiochemical 2M2B can reduce stress in transported cats based on objective physiological and behavioral measures.