AbstractThis unit provides protocols for growth of Brucella abortus on solid or in liquid media and for long‐term storage of laboratory stocks. Two issues affecting the culture and storage of isolates of this slow‐growing bacterium are emphasized: contamination of cultures and outgrowth of attenuated variants lacking a complete lipopolysaccharide. Laboratories planning to work with B. abortus should be aware that Biosafety Level 3 facilities are required. Furthermore, this organism is classified in the U.S. as a Select Agent, which restricts its use to laboratories registered with the U.S. government's Select Agent programs.