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Table 1 Proteins, source and dilution of antibodies, staining procedures and patterns and interpretation of results

From: HER2 and TOP2A in high-risk early breast cancer patients treated with adjuvant epirubicin-based dose-dense sequential chemotherapy

Protein Antibody [clone, source] Ab dilution Pretreatment/EU IT IHC Staining Detection System/Chromogen Scoring System Cut-off (%) Staining Pattern Reference
Ki67 MIB1 (1) 1:70 15'/EU2 20' Polymer HRP/DAB SQ ≥ 14%* Nuclear [28]
HER2 PL (1) 1:200 25'/EU1 30' Polymer HRP/DAB 0-3 > 30%# Membranous [23]
TopoIIa KiS1 (1) 1:200 15'/EU2 30' EnVision/DAB SQ > 5%** Nuclear [27]
ER 6F11 (2) 1:70 20'/EU1 20' Polymer HRP/DAB H-Score ≥ 1% Nuclear [29]
PgR 1A6 (2) 1:70 20'/EU1 20' Polymer HRP/DAB H-Score ≥ 1% Nuclear [29]
  1. Antigen Retrieval was done on a hot plate at 98°C.
  2. DAB: 3,3'-Diaminobenzidine; EU: Epitope Unmasking; EU1: Citric acid, pH 6.0; EU2: Ethylenediaminetetraacetate, pH 8.8; HRP: Horseradish Peroxidase; IT: Incubation time; SQ: Semiquantitative.
  3. (1): Dako, Glostrup, Denmark; (2): Leica Biosystems, Newcastle Upon Tyne, UK.
  4. *: Proliferation Index: Low if < 14% and High if ≥14% positive nuclei were observed; #: score 3+ in cases with > 30% positive tumor cells; **: positive if at least moderate staining intensity in > 5% of tumor cells.