The sustained release of antigen over a period of time is necessary to stimulate the immune system and generate a high titer of antibody. The use of adjuvants in water-in-oil emulsions has long been the favored method of prolonging the exposure time of antigens to enhance the immune response. Routine methods for preparation of this emulsion are repeated mixing of the materials in a "two-syringe" method that is labor-intensive and unreliable. The method we describe here is to use a common vortex (i.e. Vortex Genie 2, VWR) to prepare water-in-oil antigen/adjuvant emulsions. This method has been routinely used in our laboratory for many years and demonstrates its consistency with reliability for immunization of mice, hamsters, and rats and subsequently generation of hybridomas.
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