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Table 1
Antibodies and probes used
Immunohistochemistry	Manufacturer, clone	Antigen retrieval	Dilution	Evaluation

Estrogen receptor alpha	Lab Vision, SP1	0.01 M citrate NaOH, pH 6.0	1:4,000	Threshold at 5% positive nuclei
Estrogen receptor beta	Novocastra, EMR02	0.001 M EDTA-NaOH, pH 8.0	1:100	Threshold at 5% positive nuclei
Progesterone receptor	Lab Vision, SP2	0.01 M citrate NaOH, pH 6.0	1:2,000	Threshold at 5% positive nuclei
HER2/neu immunohistochemistry	Dako, polyclonal	0.01 M citrate NaOH, pH 6.0	1:200	0 - 1 + -2 + 3 + according to DAKO manual
Cytokeratin 5/6	Dako, D5/6 B4	0.001 M EDTA-NaOH, pH 8.0	1:200	Threshold at 1% positive tumor cells
Cytokeratin 14	Novocastra, LL002	0.01 M citrate NaOH, pH 6.0	1:200	Threshold at 1% positive tumor cells
Cyclin D₁immunohistochemistry	Lab Vision, SP4	0.001 M EDTA-NaOH, pH 8.0	1:800	Threshold at 5% positive nuclei
Mucin 1	Novocastra, MA695	0.001 M EDTA-NaOH, pH 8.0	1:800	0 negative; 1, 1% to 50%; 2, >50% tumor cells positive
Gross cystic fluid protein	Novocastra, 23A3	0.001 M EDTA-NaOH, pH 8.0	1:800	Threshold at 1% positive tumor cells

In situ hybridization	Probe	Protocol	Evaluation

HER2/neu amplification	Abbott PathVysion HER2 DNA Probe Kit	Manufacturer's instructions	Locus-specific signal to centromere signal ratio >2.0 considered amplified
Cyclin D₁amplification	Abbott Vysis LSI Cyclin D1/CEP11	Manufacturer's instructions	Locus-specific signal to centromere signal ratio >2.0 considered amplified

EDTA, ethylenediamine tetraacetic acid. Lab Vision (Thermo Fisher Scientific, Fremont CA, USA); Novocastra (Leica Microsystems, Wetzlar, Germany); Dako (Glostrup, Denmark); Abbott (Abbott Park, IL, USA).
Brozkova et al. Breast Cancer Research 2008 10:R48 doi:10.1186/bcr2101

Table 1