Preparation of Avidin-immune Serum

The following experiments will require (1) one 2.5 to 3.5 kg rabbit, (2) a highly purified preparation of avidin, (3) Freund s complete and incomplete adjuvants, (4) disposable Ouchterlony plates, and (5) C-biotin, which can be obtained from Amersham Searle Corp. 

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. 

During the 4 week period used to obtain control sera, determine the purity of a commercial avidin preparation using SDS-acrylamide gel electrophoresis as described in Chapter 5. Use only preparations exhibiting a single electrophoretic species in these experiments. 

The amount of avidin needed may vary, but this range of concentrations has been used successfully. 

Mix the above components by gently vortexing the solution until all components are dissolved. 

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