| Abstract |
The immunodot assay, originally described by McDougal et al., and adapted
in our laboratory, allows the determination of the heavy chain and the
light chain classes of mouse immunoglobulins (Igs). It is based on the
principle of a "sandwich immunoassay". The main feature of our assay is
the use of rat monoclonal antibodies (MAbs) anti-mouse heavy and light
chain isotype. These MAbs are dotted in sequence on strips of
nitrocellulose filter paper (5 x 80 mm, 0.22 pm porosity Schleicher and
Schuell). After saturation, the strips are incubated with the mouse
hybridoma supernatants and the mouse Igs, bound to the specific anti-mouse
isotype rat MAbs, are detected with a peroxidase-conjugated goat
polyclonal antibody directed against mouse Igs. The procedure for the test
is the same as that used for the determination of rat Ig isotypes
described in Chapter 11. The nature and the concentrations of the specific
anti-mouse Ig isotype rat MAbs are given in Table 1.
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