The liquid used for the "re-agent," as in the first film, was taken from glowsticks.
One of the original plots for the film involved Dan Cain being recruited to Washington, DC to bring the deceased President of the United States back to life with the help of Herbert West. This was later carried over to the unmade House of Re-Animator.
At one point Dr. West suggests that the noises coming from his laboratory wall are just "probably rats in the wall." This is a reference to one of H.P. Lovecraft's better known stories "Rats In The Wall."
Barbara Crampton, star of the first Re-Animator (1985) turned down the role of the Bride on advice from her agent who convinced her that it was beneath her to have such a small role. Kathleen Kinmont was cast instead. She and Crampton had worked together in a nude scene in Fraternity Vacation (1985).
A sequence was planned in the original script where the head of Dr. Hill was found as an attraction at a carnival. It is mentioned in the film but never shown, although images from that sequence are discussed on the Blu-ray.