Bond polymer refine detection system ds9800

Immunohistochemical staining in human ESCC samples was performed by using standard reagents and techniques on a Bond-Max Automated Staining System (Leica Biosystems Newcastle Ltd, Australia). The sections were incubated with primary antibodies followed by Bond Polymer Refine detection system (DS9800, Leica Biosystems, Newcastle upon Tyne, UK). The primary antibody used was p53 (clone DO7, 1:200; Leica Biosystems, Newcastle upon Tyne, UK). Positive and negative controls were performed according to manufacturer’s instruction.. The staining assessment was independently carried out by 2 pathologists (S.L.W. and W.T.H) without any information about clinicopathologic features or prognosis. The p53 expression in the tissue was scored in terms of the percentage of cells that exhibited positive nuclear staining.

p53 immunohistochemistry for the HGSC tissue samples was carried out at the Medical Research Center Oulu, Finland, using Dako (Agilent Technologies Inc., Santa Clara, CA, US) M7001 anti-human p53 antibody at 1:2400 dilution and BOND Polymer Refine Detection System DS9800 (Leica Biosystems, Deer Park, IL, US). The scoring was performed according to the WHO classification [31 ]: a strong and diffuse nuclear staining comprising > 80% of tumor cells, wild type, or total negativity in tumor cells. Both negative and excessive staining were considered pathogenic.