Immunohistochemical Analysis of ENO1 in BE and EC

Endoscopic (mucosal) biopsy specimens from BE patients and matched tumor and adjacent normal specimens from EC patients, archived as formalin-fixed, paraffin-embedded material, were used for immunohistochemical analysis. Sections were routinely stained with hematoxylin and eosin and evaluated histopathologically. Tissue specimens were processed in a standard fashion, as described elsewhere 22 (link). ENO1 was identified using an anti-ENO1 mouse monoclonal antibody at a concentration of 0.025 μg/ml (Abcam, Cambridge, UK). Immunostaining was with a peroxidase-based visualization DAKO LSAB^® kit, following the manufacturer's recommendations. Diaminobenzidine tetrahydrochloride was used as chromogen. Sections incubated with PBS instead of the anti-ENO1 antibody served as controls.