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