Bleeding, rabbits

Bleed rabbits I wk after immunization, collecting blood from the rear marginal ear vein (see Note 6). 

Rabbit Bleed (Days post initial immunization)... 

At this stage, test the antiserum using an appropriate assay (see Note 5). If the antibody has the requirements for the use to which it will be put, up to three further bleeds on successive days may be performed. If the antiserum is unsatisfactory, i.e., the reaction is very weak, inject the rabbit again 1 mo after the test bleed, and again test-bleed 10 d after this injection. 

Bleed the rabbits 10 d after the third immunization and every week thereafter. At each bleeding, check the serum as after the second immunization but the serum should be diluted at 1 4000 or 1 6000. Bleeding can be continued for as long as the antibody titer remains high (see Note 7). 

Two weeks after the second injection, test bleed the rabbit from an ear vein. Swab the ear with xylene to dilate the vein before bleeding. 

The titer of anti-RIP antibody can be determined before bleeding out the rabbit, by comparison with a sample of serum taken from the animal before immunization... 

The mixture is injected once a week for 3 weeks, and then the animal is maintained for 3 weeks without additional injections. Approximately 25-40 ml of blood is removed from the animal s ear vein to test for antibodies. One week after the first bleed the rabbit is boosted with half the antigen amount used earlier along with incomplete adjuvant incomplete adjuvant does not contain the mycobacteria. Serum is again removed 2 weeks after this injection and tested for antibody response. The enzyme-linked immunosorbent assay is used to determine if the titer (or antibody concentration) of the serum is sufficiently high to establish antibody binding to the antigen. This 3-week cycle is repeated as long as necessary to obtain the antibodies desired. 

Klement R Carlsson S, RakJ, et al. The benefit-to-risk profile of melagatran is superior to that of hirudin in a rabbit arterial thrombosis prevention and bleeding model. J Thromb Haemost 2003 1 587-594. 

Reticulocytes were obtained from rabbits made anemic by repeated bleeding. Reticulocyte counts were generally 20-80% of total red cells. Standard methods were followed for preparing and 125I-labeling of rabbit transferrin and its incubation with reticulocytes (54). Reticulocyte membranes were prepared by the method of Dodge (77) and solubilized for... 

The Simplate device can also be used to perform incisions at the shaved inner ear of rabbits taking care to avoid major vessels. The normal range of bleeding time in anaesthetized rabbits is approximately 100 s (77 4 s, n = 20). 

Klement et al. (1998) described another ear bleeding model in anaesthetized rabbits. The shaved ear was immersed in a beaker containing saline at 37 °C. Five full-thickness cuts were made with a no. 11 Bard-Parker scalpel blade avoiding major vessels and the ear was immediately re-immersed in saline. At different times thereafter (5 to 30 min) aliquots of the saline solution were removed, red cells were sedimented and lysed, and cyanohemoglobin was determined as... 

Klement P, Liao P, Hirsh J et al. (1998) Hirudin causes more bleeding than heparin in a rabbit ear bleeding model. J Lab Clin Med 132 181-185... 

The backs of 6 albino rabbits are clipped free of hair. Each material is tested on two 1-in square sites on the same animal one site is intact and one is abraded in such a way that the stratum comeum is open but no bleeding produced. Abrasion can be performed using the tip of a hypodermic needle drawn across the skin repeatedly or commercial instruments such as the... 

Since rabbits are by far the most often used animals for antibody generation, collection of their blood will be discussed here. Procedures for bleeding other small animals may be found in other references (3-5). Blood is collected from larger animals such as goats, donkeys, and horses via the jugular vein. In these cases an inexperienced investigator would be well advised to obtain the assistance of a veterinarian or other qualified individual to demonstrate proper handling of the animal and withdrawal procedures. 

Four weeks prior to injection of antigen into the rabbit, bleed the animal and process the blood as described earlier in this chapter. Repeat this procedure twice at 7 day intervals. This allows 14 days between the last bleeding and antigen injection. 

Seven days after the booster injection, bleed the rabbit and prepare cell-free serum as described in this chapter. 

Ten after the last injection, test-bleed the rabbits from the marginal... 

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