A microfluorometric assay for immunoglobulin class and subclass levels in murine serum
article
The IgG fractions of rabbit antisera specific for the Fc part of mouse IgA, IgM, and the four subclasses of IgG (1, 2a, 2b, and 3) were coupled covalently to Sepharose beads by the cyanogen bromide method. The beads were then incubated with different dilutions of mouse serum. The mouse immunoglobulins (Ig) binding to the beads were demonstrated by the use of a fluorescent goat anti mouse/Ig conjugate. Incubations were done in flat bottom microtitration plates. The fluorescence per individual bead was measured through the bottom of the plates with an inverted microfluorometer. The logarithm of the fluorescence was linearly related to the logarithm of the serum dilution for serum dilutions between 103 and 105. Ig levels of individual samples can be determined with as little as 1 μl of serum. The sensitivity of the method for IgM was estimated with MOPC 104E plasmacytoma IgM. Five ng of IgM per sample was easily detected. The method offers an attractive alternative to radioimmunoassay techniques.
Topics
TNO Identifier
228162
ISSN
0022-1759
Source
Journal of Immunological Methods, 14(3-4), pp. 213-224.
Pages
213-224
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