Formalin-fixed paraffin-embedded (FFPE) tissues were sectioned (4 µm) and forwarded to either hematoxylin and eosin (HE) or immunohistochemical staining using antibodies against brachyury (clone H-210; Santa Cruz, Santa Cruz, USA), pan-cytokeratin (clone MNF116, Dako), epithelial membrane antigen (EMA; clone E29, Dako) and S100 (polyclonal; Dako, Glostrup, Denmark). Tissue sections were subjected to antigen retrieval in a tris-borate/EDTA buffer (CC1, Ventana, Roche, Vienna, Austria) before incubating with clone H-210 (Santa Cruz, dilution 1∶50) for brachyury immunohistochemistry. Antibody detection was performed on a Ventana Immunostainer using the ultraView Universal DAB Detection Kit (Roche) according to the manufacturer’s recommendations.
El-Heliebi A., Kroneis T., Wagner K., Meditz K., Kolb D., Feichtinger J., Thallinger G.G., Quehenberger F., Liegl-Atzwanger B, & Rinner B. (2014). Resolving Tumor Heterogeneity: Genes Involved in Chordoma Cell Development Identified by Low-Template Analysis of Morphologically Distinct Cells. PLoS ONE, 9(2), e87663.
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