Comparison of the effects of microwave heating and high pressure cooking for antigen retrieval of human and rat Bcl-2 protein in formaldehyde-fixed, paraffin-embedded sections
| Autor: | H-J van Slooten, Rob W.M. Hoetelmans, C.J.H. van de Velde, R. Keijzer, J. H. Van Dierendonck |
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| Rok vydání: | 2002 |
| Předmět: | |
| Zdroj: | Biotechnic & Histochemistry. 77:137-144 |
| ISSN: | 1473-7760 1052-0295 |
| DOI: | 10.1080/bih.77.3.137.144 |
| Popis: | Immunohistochemical detection of expression of the anti-apoptotic Bcl-2 protein is widely studied as a putative prognostic and predictive factor in various types of cancer. For that purpose, heating for 10 min by microwave (MW) up to 100o C in citrate buffer, pH 6.0, prior to immunostaining is often used to retrieve Bcl-2 antigens in archival formalin-fixed, paraffin-embedded tissue. We recently reported that Bcl-2 is not only a cytoplasmic protein, but that it is present also in interphase nuclei and that it strongly associates with mitotic chromosomes. Furthermore, we showed that binding of the monoclonal antibody (MAb) #124 with nuclear/chromosomal epitopes is diminished by formaldehyde-based fixatives and cannot be restored by MW treatment for 10 min. Here we report that prolonged MW heating or heating up to 130°C in a high pressure cooker (HPC), despite improved cytoplasmic immunostaining, fails to retrieve nuclear/chromosomal Bcl-2 epitopes recognized by the MAb #124 in human tissues. In contrast, t... |
| Databáze: | OpenAIRE |
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