PMID: 6969247May 1, 1980Paper

Immunoelectron microscopy of cell surface antigens: a quantitative analysis of antibody binding after different fixation protocols

The Histochemical Journal
W Van EwijkI L Weissman

Abstract

The effect of different fixation solutions on the denaturation of membrane-associated antigens in murine lymphoid cells was determined quantitatively using microfluorometric analysis and a radioimmunoassay. Paraformaldehyde and periodate-lysine-paraformaldehyde solutions preserved the antigenicity of cell surface-associated immunoglobulin (S-Ig) antigens when used in concentrations ranging from 0.01 to 4%. However, glutaraldehyde destroyed the antigenicity of S-Ig and Thy 1.2 molecules at concentrations higher than 0.1%. Electron microscopic analysis of the different fixed cell suspensions, after labelling of the cells with a rabbit anti-mouse immunoglobulin-horseradish peroxidase conjugate (RaM-Ig-HRP) showed that prefixation of the sample with 0.1% glutaraldehyde was optimal for immunoelectron microscopical studies, since this concentration preserved both the antigenicity of membrane-associated antigens as well as the ultrastructure of the cells under study. Prolonged fixation periods affected antibody binding. However, S-Ig molecules denatured at a slower rate than Thy 1.2 molecules. A preparation method for the immunoelectron microscopical localization of lymphoid and non-lymphoid cell types in lymphoid organs is reported.

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Citations

Jan 1, 1988·Rheumatology International·P I Mapp, P A Revell
Sep 1, 1982·The Histochemical Journal·M A Judd, K J Britten
Feb 1, 1984·The Histochemical Journal·W Van EwijkJ F Jongkind
Apr 15, 1997·Proceedings of the National Academy of Sciences of the United States of America·W Van EwijkT Logtenberg
Sep 1, 1991·The Histochemical Journal·J P De JongR E Ploemacher
Jan 1, 1984·Virchows Archiv. B, Cell Pathology Including Molecular Pathology·M L HansmannM R Parwaresch
May 1, 1984·Immunobiology·A A GeldofH L Langevoort
May 1, 1982·Journal of the American Academy of Dermatology·E M McMillanM A Everett
Dec 1, 1983·Journal of Cutaneous Pathology·E M McMillan
Jun 1, 1981·Journal of Cutaneous Pathology·E M McMillanM A Everett
Jul 1, 1988·European Journal of Haematology·H E HeierE Falleth
Dec 17, 1985·Journal of Immunological Methods·N R SwansonR A Joske

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