Table 1.
Staining details Ki67, CD31, Ang2/Tie2, and CD31+CD34/aSMA, CD4, CD8, and CD16.
| Staining performed at HistogeneX | ||
|---|---|---|
| Antibody clone | Detection system | Staining instrument |
| CD4 – clone 1F6 monoclonal mouse Ab (Novocastra - NCL-CD4-1F6) |
Optiview DAB | Ventana Benchmark XT |
| CD8 – clone C8/144B (Dako, M7103) monoclonal mouse Ab | Ultraview DAB | Ventana Benchmark XT |
| CD16 – clone SP175 monoclonal rabbit Ab (Ventana – 760-4863) | Ultraview DAB | Ventana Benchmark XT |
| CD31 – clone 1A10 monoclonal mouse Ab (Novocastra – NCL-CD31-1A10) Ki67 – clone 30.9 monoclonal rabbit Ab (Ventana – 790-4286) |
Cy3 – goat anti-mouse (Jackson ImmunoResearch 115–165–166) Alexa488 – goat anti-rabbit (Invitrogen, A11070) |
Ventana Discovery XT |
| CD31 – clone 1A10 monoclonal mouse Ab (Novocastra – NCL-CD31-1A10) CD34 clone QBEnd/10 monoclonal mouse Ab (Ventana, 790-2927) aSMA – polyclonal rabbit Ab (Abcam, ab5694) |
Alexa647 – goat anti-mouse (Invitrogen A21236) Alexa647 – goat anti-mouse (Invitrogen A21236) Alexa488 – goat anti-rabbit (Molecular probes A11070) |
Ventana Discovery XT |
| Ang2 – clone F-1 monoclonal mouse Ab (Santa Cruz Technology, sc-74,403) Tie2 – polyclonal goat Ab (R&D Systems, AF313) |
Cy3 – rabbit anti-mouse (Invitrogen A11078) Alexa488 – rabbit anti-goat (Jackson Immunoresearch 315-165-003) |
Ventana Discovery XT |
| Staining performed in the Good Clinical Practice Pathology Laboratory in Penzberg | ||
| CD3 – clone 2GV6 monoclonal rabbit Ab (Ventana – 790-4341) |
Ultraview AP Red | Ventana Benchmark Ultra |
| CD68 – clone KP1 monoclonal mouse Ab (Ventana – 760-2931) CD163 – clone MRQ-26 monoclonal mouse Ab (Cell Marque), ready to use Dispenser (Ventana – 760-4437) |
Ultraview AP Red (Ventana – 760-501) Optiview DAB (Ventana – 760-700) |
Ventana Benchmark XT Ventana Benchmark XT |
Ab, antibody; CD, cluster of differentiation.