Notes

1. Balb-c mice are an inbred strain ideally suited to monoclonal antibody work. Females are usually used because they do not fight when housed together in project groups of three to five individuals. This mouse is also known as the "barber" strain because the dominant female will remove the whiskers from the others in the group.

2. New Zealand rabbits are normally used for serum production; they are easily handled and adapt well to individual cages or group floor pens. This strain has half-lop ears, which make blood collection from marginal veins a fairly straightforward procedure.

3. Most antigens require an adjuvant to increase their immunogenicity and a number of formulations can be used. Regulations on their use should be consulted prior to embarking on a course of immunizations. For many years Freund's complete and incomplete adjuvant were the formulation of choice for all immunization work. In recent years welfare issues have been raised over the use of these adjuvants and a number of alternatives based on water-soluble bacterial cell wall components have become available.

4. Parenteral anesthetic agents are preferable to gaseous ones as the size of the mouse creates problems when using standard anesthetic machines.

5. Tail tip amputation is normally used to obtain test bleeds from mice, and this is normally carried out under light anesthesia induced with diethyl ether.

6. It is extremely important that the medium is harvested after 24 h of incubation. Longer incubation periods may induce the formation of suppressing cytokines, which will block the desired cell stimulation.

7. It is important that the thymocytes are not disturbed for the full 72-h incubation and that they are rapidly harvested and fused after this period of time.

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