EGFR IHC scores and test details for tissue samples in each laboratory
| Tissue sample | Laboratory | |||||||
| A | B | C | D | E | F | G | H | |
| 1 | 0 | 0 | 0 | 1 | 3 | 3 | 1 | 1 |
| 2 | 1 | 1 | 1 | 2 | 3 | 3 | 1 | 2 |
| 3 | 1 | 1 | 0 | 0 | 1 | 1 | 1 | 2 |
| 4 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 5 | 1 | 1 | 0 | 0 | 2 | 2 | 1 | 1 |
| 6 | 1 | 0 | 0 | 0 | 0 | 1 | 0 | 0 |
| 7 | 2 | 1 | 0 | 0 | 3 | 3 | 2 | 3 |
| 8 | 2 | 1 | 0 | 1 | 3 | 3 | 1 | 1 |
| 9 | 0 | 0 | 0 | 0 | 1 | 2 | 1 | 0 |
| 10 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
| 11 | 1 | 1 | 0 | 1 | 2 | 2 | 1 | 1 |
| 12 | 2 | 3 | 1 | 2 | 3 | 3 | 2 | 3 |
| 13 | 3 | 2 | 2 | 2 | 3 | 3 | 2 | 2 |
| 14 | 3 | 1 | 0 | 1 | 3 | 3 | 1 | 2 |
| 15 | 2 | 1 | 0 | 0 | 2 | 3 | 1 | 0 |
| 16 | 1 | 0 | 0 | 0 | 1 | 1 | 0 | 0 |
| 17 | 3 | 1 | 2 | 1 | 1 | 3 | 3 | 0 |
| Antibody | EGFR.113* | Clone 31G7‡ | Clone 111.6† | Clone 111.6† | Clone 31G7‡ | Clone EGFR.25* | Clone 30§ | Clone 2-18c9§ |
| Epitope | Extracellular | Extracellular | Extracellular | Extracellular | Extracellular | Cytoplasm domain | Extracellular | Extracellular |
| Dilution | 1:125, 45 min | 1:40 | 1:50 | 1:400, 60 min RT | 1:200, o/n 4'C | 1:25 | 1:25 | $ |
| Epitope retrieval | 10 min Tris-EDTA pH 9 | Boiling DM809 buffer*** pH 6.1 10 min | Protease 2, 32 min‡‡ | Protease type XIV,0.05%, 5 min | Protease, 0.1 M, 15 min RT | Tris-EDTA pH 8.8, ER2** | Protease 8 min¶ | Prot K 5 in RT |
| Staining | PowerVision, 30 min RT‡‡ | Envision DM802§ | †† | PowerVision¶¶ | PowerVision | PowerVision§§ | $ | |
The number and content of each tissue microarray sample is as described in the legend for Table 3. The laboratory codes do not correspond to those in Table 3.
Lab A uses 1) Poseidon Amsterdam, NL; Lab B uses 2) Vysis, Hoofddorp, NL; Lab C uses 2) Vysis, Hoofddorp, NL; Lab D uses 2) Vysis, Hoofddorp, NL; Lab E uses 3) Zytovision, Immunologic, Duiven NL; Lab F uses 3) Zytovision, Immunologic, Duiven NL; Lab G uses 3) Zytovision, Immunologic, Duiven NL; Lab H uses 4) DAKO, Glostrup, DK.
↵* Novacastra, Newcastle, UK.
↵† NeoMarkers; Stratech Scientific, Newmarket, UK.
↵‡ Zymed.
↵§ Dako, Glostrup, Denmark
↵¶ Ventana prepkit.
↵** Bond-maX.
↵†† Ventana Benchmark XT.
↵‡‡ nr pdvo-hrp 110.
↵§§ Vision biosystems Bond-maX.
↵¶¶ Immunologic via klinipath.
↵$ Pharm Dx IHC Kit
EGFR, epidermal growth factor receptor; IHC, immunohistochemistry; o/n, overnight.
RT = Room Temperature