In rats and mice, only one footpad may be used. The concomitant use of appropriate analgesics is required if footpad injections are used. Animals should be maintained on soft bedding and not on wire-bottomed cages. The skillful preparation and delivery of the antigen is critical to the success of the immunization and comfort of the animal. If necessary animals, especially rabbits, may be sedated prior to injection of Freunds Adjuvant.
Improper delivery results in tissue necrosis and failure.
The formulation strength of CFA is important in determining the degree of reaction and side effects. If an abscess or ulceration develops at any injection site, it must be reported through established channels, e.g., the animal resources supervisor or veterinarian, and must receive appropriate veterinary treatment. CCAC Guidelines on Acceptable Immunological Procedures . The volume must not exceed 0.1 ml. In priming prior to injecting hybridomas, it appears that 0.1 ml is an adequate dose of pristane.