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A Simple Epitope Retrieval Method Without the Use of Microwave Oven or Enzyme Digestion

A Simple Epitope Retrieval Method Without the Use of Microwave Oven or Enzyme Digestion A simple epitope retrieval method has been developed in our research laboratory for immunostaining of formalin-fixed, paraffin-embedded tissues. This method is easy to use, as it only involves an overnight incubation of deparaffinized sections in a regular oven at 70 to 80°C with 10m Mcitrate buffer (pH 6.0) prior to immunostaining. With this approach, a variety of nuclear and cytoplasmic antigens known to require microwave irradiation or enzymatic digestion for their optimal detection could be clearly demonstrated with excellent preservation of morphology, intense reactivities, and clean backgrounds. Also, optimal immunostaining for each of the 15 antibodies tested was observed even when sections from 15 different cases were incubated together with 10m Mcitrate buffer, then immunostained in an identical manner with manufacturers' recommended antibody concentrations. The drawback of our method is that it takes longer to yield results. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Immunohistochemistry & Molecular Morphology Wolters Kluwer Health

A Simple Epitope Retrieval Method Without the Use of Microwave Oven or Enzyme Digestion

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ISSN
1062-3345
eISSN
1533-4058

Abstract

A simple epitope retrieval method has been developed in our research laboratory for immunostaining of formalin-fixed, paraffin-embedded tissues. This method is easy to use, as it only involves an overnight incubation of deparaffinized sections in a regular oven at 70 to 80°C with 10m Mcitrate buffer (pH 6.0) prior to immunostaining. With this approach, a variety of nuclear and cytoplasmic antigens known to require microwave irradiation or enzymatic digestion for their optimal detection could be clearly demonstrated with excellent preservation of morphology, intense reactivities, and clean backgrounds. Also, optimal immunostaining for each of the 15 antibodies tested was observed even when sections from 15 different cases were incubated together with 10m Mcitrate buffer, then immunostained in an identical manner with manufacturers' recommended antibody concentrations. The drawback of our method is that it takes longer to yield results.

Journal

Applied Immunohistochemistry & Molecular Morphology – Wolters Kluwer Health

Published: Jan 1, 1996

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