|MadSci Network: Immunology|
Probably the best way to bias toward IgG production is to hyperimmunize subcutaneously and to use draining lymph nodes rather than spleens for fusion. The first injection (50-100 microliters) should be in Complete Freund’s Adjuvant subcutaneously near the hind feet. Four subsequent injections (50-100 ul) are given subq. in PBS. Fuse one day after the last injection. Do not use peak serum antibody titers as a guide for when to fuse – studies have shown that dividing cells fuse preferentially. The antibody-secreting plasma cells are non-dividing. Dissect the draining lymph nodes, which will be fairly large because of the CFA stimulation.
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