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Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Stumptner, C; Gogg-Kamerer, M; Viertler, C; Denk, H; Zatloukal, K.
Immunofluorescence and Immunohistochemical Detection of Keratins.
Methods Enzymol. 2016; 568:139-162
Doi: 10.1016/bs.mie.2015.08.003
Web of Science
PubMed
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FullText_MUG
- Führende Autor*innen der Med Uni Graz
- Stumptner Cornelia
- Zatloukal Kurt
- Co-Autor*innen der Med Uni Graz
- Gogg-Kamerer Margit
- Viertler Christian
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- Abstract:
- Reliable detection of keratins in tissues is important for investigating their physiological role and for using keratin expression as a biomarker in medical diagnostics. A particular challenge for the detection of keratins by immunofluorescence microscopy or immunohistochemistry relates to the fact that keratin intermediate filaments are obligatory heteropolymers, which may result in dissociation between RNA and protein expression levels in the event that the homeostasis of the expression of the proper keratin partners is disturbed. Furthermore, variable accessibility of epitopes on keratin polypeptides due to conformational changes may lead to false negative results. Preanalytical effects, such as warm/cold ischemia, fixation, tissue processing, and embedding may result in false negative or inappropriate reactions. An experimental design for how to systematically test preanalytical effects and to validate immunohistochemistry protocols is presented. This kind of evaluation should be performed for each antigen and antibody since the various epitopes recognized by antibodies may behave differently. In this context, one has to be aware that different cell structures may be affected or modified differently by various preanalytical procedures and may thus require different preanalytical and staining protocols. Copyright © 2016 Elsevier Inc. All rights reserved.