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Selective depletion of immunoglobulin isotypes after decomplementation of mouse sera by heat treatment
T Schetters1, J Van Run-Van Breda, A Van Zon
1Department of Cytology and Histology, University of Nijmegen, The Netherlands.
Inactivation of mouse serum by heat treatment (30 min at 56 degrees C) decreased antibody reactivity as shown by haemagglutination assays. This appeared to be due to loss of antibody since Ouchterlony analysis showed disappearance of antibody after heat treatment. By both isotype-specific ELISA and Ouchterlony analysis it was shown that mouse IgM, IgG2b and IgG3 are sensitive to heat treatment whereas IgG2a is relatively resistant. IgG1 is not sensitive to heat treatment. To avoid complement-dependent lysis in haemagglutination assays EDTA should be added to the serum samples.
Inactivation of mouse serum by heat treatment (30 min at 56 degrees C) decreased antibody reactivity as shown by haemagglutination assays. This appeared to be due to loss of antibody since Ouchterlony analysis showed disappearance of antibody after heat treatment. By both isotype-specific ELISA and Ouchterlony analysis it was shown that mouse IgM, IgG2b and IgG3 are sensitive to heat treatment whereas IgG2a is relatively resistant. IgG1 is not sensitive to heat treatment. To avoid complement-dependent lysis in haemagglutination assays EDTA should be added to the serum samples.