Anti epcam clone ber ep4

HCC specimens collected from patients who had undergone liver biopsy were formalin-fixed and paraffin-embedded (FFPE). In each HCC specimen, the histological subtypes [22 (link)] and grades were determined according to the WHO classification by an expert liver pathologist (B.R.).
The following primary antibodies were used: anti-EpCAM (clone Ber-EP4, Dako, Agilent technology France, Les Ulis, France), anti-CK19 (clone RCK108, Dako, Agilent technology France, Les Ulis, France), anti-glutamine synthetase (GS, clone 6/GS, BD Biosciences, Le Pont de Claix, France) and anti-p53 (clone DO-7, Roche/Ventana). Immunohistochemical assays were performed on the Ventana Benchmark Ultra automated staining instrument (Ventana Medical Systems, Illkirch, France). For each antibody, staining was analysed using the following scores: surface (0: 0–5%, 1: 5–25%, 2: 25–50% and 3: >50%) and intensity (0: no expression, 1: weak expression and 2: high expression). Stainings were considered positive if the percentage of tumor cells was >5%.

Sections of 4 µm were cut from formalin-fixed and paraffin-embedded (FFPE) blocks of cell lines and incubated with anti-Aurora A (HPA002636, Sigma, Darmstadt, Germany, dilution 1: 250), anti-Bcl-xL (2762S, Cell Signaling Technology, Frankfurt, Germany, dilution 1: 1000), anti-vimentin (clone V9, Dako, Glostrup, Denmark, dilution 1:8000) and anti-EPCAM (clone Ber-EP4, Dako, Glostrup, Denmark, dilution 1:250) antibodies. Microscopy of the cells was performed on a Leica DMLB microscope (1003/numerical aperture [NA] 1.25; Leica Microsystems, Wetzlar, Germany) with 40× objectives using LasV3.6 acquisition software.