Materials and Methods
Sources of Animals
The source and maintenance of bluegill are described in Chapter
II. Mice used as comparative controls were obtained from an outbred
albino strain and were kindly provided from a colony being maintained
by Dr. George Gifford, University of Florida.
Sources of Antigens and Antisera
Rabbit antisera to grouper (a marine teleost fish) light (L) chains
(Ra-GL chain) were the same as those used previously (29). Rabbit anti
sera to bluegill serum Ig were prepared by immunizing rabbits with im
mune precipitates of bream serum proteins precipitated at equivalence
with Ra-GL chain. This approach took advantage of the considerable
cross reactivity between L chains of different species of fish (L. W.
%
Clem, unpublished observations). Each of three rabbits received three
biweekly subcutaneous injections of immune precipitate (1 mg/injection)
in complete Freunds adjuvant. The antisera obtained by bleeding at 7-
9 weeks were pooled and used here. The specificity of the rabbit anti-
bluegill immunoglobulin (Ra-Blg) is discussed under Results. Sheep
antiserum to rabbit IgG was prepared by repeated (weekly for four
weeks) injections of alum precipitated rabbit IgG (purified by DEAE-
cellulose chromatography from Fraction II, Pentex, Kankakee, Ill.). The
antiserum obtained at 5-6 weeks was specific for rabbit IgG and contained
130