Immunohistochemical Labeling of Breast Cancer Markers
Immunohistochemical labeling was performed according to standard protocols on serial 2.5 μm thick sections from the original blocks or the TMA blocks. To assure optimal reactivity, immunostaining was applied 7–10 days after sectioning at the Laboratory of Molecular Oncology of the Hellenic Foundation for Cancer Research, Aristotle University of Thessaloniki School of Medicine. The staining procedures for HER2 (A0485 polyclonal antibody, Dako, Glostrup, Denmark), estrogen receptor (ER, clone 6F11, Novocastra™, Leica Biosystems, Newcastle, U.K), progesterone receptor (PgR, clone 1A6, Novocastra™, Leica Biosystems) and Ki67 (clone MIB-1, Dako) were performed using a Bond Max™ autostainer (Leica Microsystems, Wetzlar, Germany), as previously described in detail [25 (link)–28 (link)]. The staining procedures for OPN (clone OP3 N, code NCL-O-PONTIN, Novocastra™, Leica Biosystems) were also performed using a Bond Max™ autostainer, as previously described [29 (link)].
Psyrri A., Kalogeras K.T., Wirtz R.M., Kouvatseas G., Karayannopoulou G., Goussia A., Zagouri F., Veltrup E., Timotheadou E., Gogas H., Koutras A., Lazaridis G., Christodoulou C., Pentheroudakis G., Economopoulou P., Laskarakis A., Arapantoni-Dadioti P., Batistatou A., Sotiropoulou M., Aravantinos G., Papakostas P., Kosmidis P., Pectasides D, & Fountzilas G. (2017). Association of osteopontin with specific prognostic factors and survival in adjuvant breast cancer trials of the Hellenic Cooperative Oncology Group. Journal of Translational Medicine, 15, 30.
Other organizations :
University General Hospital Attikon, Aristotle University of Thessaloniki, University Hospital of Ioannina, Alexandra Hospital, National and Kapodistrian University of Athens, Papageorgiou General Hospital, Laiko General Hospital of Athens, University of Patras, Metropolitan Hospital, Hospital Agioi Anargyroi, Hippocration General Hospital, Hygeia Hospital
Staining procedures for HER2, ER, PgR, Ki67, and OPN
dependent variables
Protein expression levels of HER2, ER, PgR, Ki67, and OPN
control variables
Serial 2.5 μm thick sections from the original blocks or the TMA blocks
Optimal reactivity achieved by applying immunostaining 7–10 days after sectioning
Staining procedures performed using a Bond Max™ autostainer
controls
Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned
Annotations
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