Whenever possible, in vitro methods should be used first to determine whether acceptable antibody titers may be achieved. Ascitic fluids may be contaminated with rodent plasma proteins, immunoglobulins, cytokines, and infectious agents. Is ascites production is required, priming procedures should be fully described. Most procedures call for 0.1-0.2 ml pristane or IFA injected intraperitoneally 2 weeks prior to hybridoma inoculation. Following injection of hybridoma cells, mice should be monitored daily for the production of ascites fluid.
Isofluorane sedation is recommended.
Replacement of a portion of the fluid volume with 1-2 ml of warm physiological saline may decrease adverse effects. Mice should be monitored for a minimum of 30 min after each tap to detect signs of circulatory shock. Mice should be tapped a maximum of three times, with the third time post-euthanasia. Mice should be euthanized anytime they show signs of labored breathing, difficulty in movement, or other signs of distress. Numbers of mice to be used for ascites production should be justified.