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Immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH) analysis of MET and KRAS. For IHC, the specific MET antibody (Met (D1C2) XP Rabbit.

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Presentation on theme: "Immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH) analysis of MET and KRAS. For IHC, the specific MET antibody (Met (D1C2) XP Rabbit."— Presentation transcript:

1 Immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH) analysis of MET and KRAS. For IHC, the specific MET antibody (Met (D1C2) XP Rabbit mAB, Cell Signaling Technology, Inc.; dilution 1:1000) and KRAS (F234) antibody (SC-30, mouse monoclonal IgG2a Santa Cruz Biotechnology; dilution 1:100) have been used. Immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH) analysis of MET and KRAS. For IHC, the specific MET antibody (Met (D1C2) XP Rabbit mAB, Cell Signaling Technology, Inc.; dilution 1:1000) and KRAS (F234) antibody (SC-30, mouse monoclonal IgG2a Santa Cruz Biotechnology; dilution 1:100) have been used. For FISH analysis the c-MET amplification probe (cytocell) and KRAS/CEN12q FISH probe (Abnova) have been used. Magnification for IHC pictures is ×200 (scale bar: 100 µm), except for B2 that have ×40 (scale bar: 500 µm). Magnification for FISH pictures is ×630 (scale bar: 10 µm). (A) Liver metastasis. (1) IHC showing cytoplasmic MET overexpression (red staining); (2) MET gene amplification (red dots) by FISH in tumour nuclei; (3) IHC negative staining for KRAS protein expression in the same tumour area where MET protein is overexpressed. (B) Ureteral metastasis. (1) IHC showing cytoplasmic MET overexpression (red staining) and FISH analysis (inset) showing MET gene amplification; (2) dual-IHC assay showing overexpression of MET (red staining, upper left) and KRAS (brown staining, bottom right) proteins in two different areas of the same specimen; (3) showing cytoplasmic and membrane KRAS overexpression (red staining) and FISH analysis (inset) showing KRAS gene amplification in the tumour nuclei. (C) Retroperitoneal metastasis. (1) IHC analysis showing negative staining for MET protein; (2) FISH analysis showing KRAS gene amplification (red dots) in tumour nuclei; (3) IHC showing cytoplasmic and membrane KRAS overexpression (red staining) in the same area where MET staining was negative. Andrea Sartore-Bianchi et al. ESMO Open 2016;1:e000079 Copyright © European Society for Medical Oncology. All rights reserved.

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