Table 2 Characteristics for anti-HER and anti-Ki-67 immunostaining and scoring
From: Cytogenetic analysis of HER1/EGFR, HER2, HER3 and HER4 in 278 breast cancer patients
Characteristic | HER1 | HER2 | HER3 | HER4 | Ki-67 |
|---|---|---|---|---|---|
Antibody | Mouse mAb | Rabbit pAb | Mouse mAb | Rabbit mAb | Mouse mAb |
Origin | Dako | Dako | NanoTools | Cell Signaling | Dako |
Clone | 2–18C9 | - | 5A12 | 83B10 | MIB-1 |
Concentration of primary antibody | EGFR pharmDX | HercepTest (2.5 μg/ml) | 5 μg/ml culture supernatant | 0.7 μg/ml | 1.6 μg/ml |
Staining pattern | Membrane | Membrane | Cytoplasm and membrane | Cytoplasm and membrane | Nucleus |
Epitope retrieval | Proteinase K | Heat induced, 10 mM citric acid buffer, pH 6.0 | Heat induced, 10 mM citric acid buffer, pH 7.3 | Heat induced, 10 mM sodium-citric acid pH 6.0 | Heat induced, 10 mM citric acid buffer, pH 7.2 |
Blocking | Endogenous peroxidase blocking | Endogenous peroxidase blocking | Endogenous peroxidase blocking | Endogenous peroxidase blocking | Endogenous peroxidase blocking |
Primary antibody | Overnight, 4°C | Overnight, 4°C | Overnight, 4°C | Overnight, 4°C | 30 min, room temperature |
Detection system | EnVisionâ„¢ Dual Link System (Dako); DAB + chromogenic substrate | EnVisionâ„¢ Dual Link System (Dako); DAB + chromogenic substrate | EnVisionâ„¢ Dual Link System (Dako); DAB + chromogenic substrate | EnVisionâ„¢ Dual Link System (Dako); DAB + chromogenic substrate | iVIEWâ„¢ DAB Detection Kit (Ventana) |
Scoring in accordance with | EGFR pharmDx guidelines | HercepTest guidelines | EGFR pharmDx guidelines | EGFR pharmDx guidelines | Manufacturer's guidelines |