A simple method for the preparation of antisera specific for murine immunoglobulin heavy chains

Rabbit antisera were prepared against mouse immunoglobulin mu and gamma chains by immunizing rabbits with their own erythrocytes coated with murine anti-rabbit erythrocyte immunoglobulin. In the preparation of these antisera, we exploited the observation that the primary immune response in C57BL/10ScSn (B10) mice is almost exclusively IgM, while hyperimmune antiserum of A/WySn (A) is mostly IgG. After appropriate absorption with thymocytes and rabbit erythrocytes (Rrbc) coated with B10 or A antiRrbc Ig, the sera were shown to be specific by immunoelectrophoresis, indirect immunofluorescent staining, and complement-mediated cytotoxicity. This technique is simple, requires almost no equipment, and a large quantity of specific antiserum can be prepared quite economically.