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Epitope Retrieval with Microwaves

Epitope Retrieval with Microwaves This study compares the efficacy of three commercial epitope retrieval solutions and two “homemade” reagents, namely, citrate buffer solution at pH 6.0 and EDTA at pH 8.0, in the restoration of immunoreactivity in a variety of nonlesional and tumor tissues. Immuno-staining was performed using 85 common diagnostic antibodies, Dako target retrieval solution (Dako TRS) produced the best overall results compared with Monosan target unmasking fluid, Serotec target unmasking fluid, EDTA and citrate buffer solution, the last four producing somewhat similar results. In addition to noticeable improvement in the intensity of staining for antibodies such as estrogen receptor, progesterone receptor, Factor VIII-related protein, surfactant, and some hematolymphoid antibodies (such as CD43, VS38C, and CD35), Dako TRS made it possible to demonstrate the CD5 antigen with Leul, an antibody not been previously reported to be immunoreactive in fixed, paraffin-embedded sections. Dako TRS was the cheapest of the three commercial reagents and appears to be reusable for as many as 20 times without a detectable loss in reactivity. Because of costs, this reagent may be best employed for the restricted number of antibodies that showed greatest benefit from its use, with homemade citrate buffer and EDTA producing equally good results in most instances. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Immunohistochemistry & Molecular Morphology Wolters Kluwer Health

Epitope Retrieval with Microwaves

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ISSN
1062-3345
eISSN
1533-4058

Abstract

This study compares the efficacy of three commercial epitope retrieval solutions and two “homemade” reagents, namely, citrate buffer solution at pH 6.0 and EDTA at pH 8.0, in the restoration of immunoreactivity in a variety of nonlesional and tumor tissues. Immuno-staining was performed using 85 common diagnostic antibodies, Dako target retrieval solution (Dako TRS) produced the best overall results compared with Monosan target unmasking fluid, Serotec target unmasking fluid, EDTA and citrate buffer solution, the last four producing somewhat similar results. In addition to noticeable improvement in the intensity of staining for antibodies such as estrogen receptor, progesterone receptor, Factor VIII-related protein, surfactant, and some hematolymphoid antibodies (such as CD43, VS38C, and CD35), Dako TRS made it possible to demonstrate the CD5 antigen with Leul, an antibody not been previously reported to be immunoreactive in fixed, paraffin-embedded sections. Dako TRS was the cheapest of the three commercial reagents and appears to be reusable for as many as 20 times without a detectable loss in reactivity. Because of costs, this reagent may be best employed for the restricted number of antibodies that showed greatest benefit from its use, with homemade citrate buffer and EDTA producing equally good results in most instances.

Journal

Applied Immunohistochemistry & Molecular Morphology – Wolters Kluwer Health

Published: Jan 1, 1996

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