Application of the critical molar concentration concept to heat-mediated antigen retrieval in immunohistochemistry |
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Authors: | Montero C Chi-Ahumada E Chavez-Porras S Gutierrez M |
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Institution: | (1) Department of Pathology, Hospital Central Dr. Ignacio Morones Prieto , Facultad de Medicina, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México;(2) Department of Morphological Sciences, Faculty of Medicine, University of Cádiz, Cádiz, Spain |
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Abstract: | Previously we have applied Scott's critical molar concentration concept to show that divalent cations, especially Mg2+ may be used to measure the affinity of a known monoclonal antibody for its antigen. In this paper we report the application of this same procedure to a study of a series of antigens (three globular proteins and three intermediate filaments). The concept was applied to samples without any previous treatment or after the application of heat-mediated antigen retrieval (using a pressure cooker). Our findings suggest that heat-mediated antigen retrieval sets free protein side-chain(s) that have been masked by formaldehyde fixation. This is reflected in a higher affinity of the antibody for the antigen in question. |
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