PMID: 2495321May 1, 1989Paper

Antigen preservation tests for immunocytochemical detection of cytoskeletal proteins: influence of aldehyde fixatives

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
B M Riederer

Abstract

The effects of aldehyde fixatives on immunochemical detection of cytoskeletal proteins were demonstrated by applying several quantitative assays to evaluate antigen conservation. Immunologically detectable brain spectrin (240/235) was measured by dot-immunobinding and quantitative immunodot assay using a polyclonal antibody. Paraformaldehyde fixation led to a 43-66% reduction in brain spectrin (240/235) immunodetection, and increasing glutaraldehyde concentrations decreased the immunological detection even more. Quantitative cryosection immunoassay and immunocytochemical localization confirmed the aldehyde sensitivity of brain spectrin (240/235). Brain spectrin (240/235) immunoreactivity decreased with increasing protein crosslinking and was dependent on glutaraldehyde concentration and post-fixation period. The assays were also used to test for conservation of antigenicity of neurofilament proteins by two monoclonal antibodies. Neurofilament detection was abolished in brain tissue after aldehyde fixation. The described methods allow screening within 24 hr of many fixation conditions by use of purified proteins as well as brain tissue samples, and allow an estimate of fixative influence on the conservation of protein antigenicity.

Citations

Jul 1, 1989·Brain Research Bulletin·I S Zagon, S R Goodman
Dec 17, 2008·Forensic Science International. Genetics·Richard ThorogateBarbara Daniel
Oct 24, 2006·Journal of Immunological Methods·Kodela VaniSeshi R Sompuram
Aug 28, 2010·International Journal of Alzheimer's Disease·Kwang-Jin OhElliott J Mufson
Dec 6, 2005·Journal of Immunological Methods·Julie St-LaurentLouis-Philippe Boulet
Aug 21, 2007·Experimental Neurology·Glenda M Halliday, Heather McCann
Jul 1, 1990·Arteriosclerosis : an Official Journal of the American Heart Association, Inc·B Harrach, H Robenek
Jan 1, 1993·Arteriosclerosis and Thrombosis : a Journal of Vascular Biology·B KaesbergH Robenek
Jan 22, 1996·The Journal of Comparative Neurology·B M RiedererR A Marugg

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