The use of monoclonal antibodies with colloidal gold-labeled probes in postembedding immunoelectron microscopy |
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Authors: | Carlos E Suarez Ruth Brown |
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Institution: | (1) Department of Veterinary Microbiology and Pathology, Washington State University, 99164-7040 Pullman, WA;(2) Washington Disease Diagnostic Laboratory, Washington State University, 99164-7040 Pullman, WA |
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Abstract: | This article focuses on procedures for the use of monoclonal antibodies (MAb) in immunoelectron microscopy (IEM) for the subcellular
localization of antigens or to relate function with structure in prokaryotic and eukaryotic cells using postembedding immunoelectronmicroscopy
techniques.
The use of MAbs greatly increases the specificity and quality of information when used in combination with gold-labeled probes.
Because of its specificity, the reactivity of MAb may be very sensitive to antigenic changes resulting from the process of
sample preparation when performing IEM studies. Specific protocols for each particular combination of epitope/MAb must be
usually specifically devised, since it is impossible to predict an experimental system that will successfully preserve structures
and antigenic determinants for every combination. In this article, we discuss critical technical aspects that usually result
in improved resolution when the procedure is used to identify structures in diverse prokaryotic, and eukaryotic cells. |
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Keywords: | Monoclonal antibodies immunoelectronmicroscopy epitope localization |
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