MSH2 Polyclonal antibody

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Tumor specimens were fixed in 10% neutral-buffered formalin for 48 h, embedded in paraffin, and cut into 4-μm-thick sections, and then mounted on silane-coated glass slides. Antigen-retrieval was done by autoclaving (121 °C) for 5 min in pH 9 Antigen Retrieval Liquid (Nichirei, Tokyo, Japan) for MUC2, MUC5AC, and hMSH2, and by microwave irradiation for 10 min in pH 9 Antigen Retrieval Liquid for human mutL homolog 1 (hMLH1). Primary antibodies used were the mouse monoclonal antibody for MUC2 (1:100 dilution; clone Ccp58, Novocastra, Newcastle Upon Tyne, United Kingdom), the mouse monoclonal antibody for MUC5AC (1:100 dilution; CLH2, Novocastra), the rabbit anti-MLH1 monoclonal antibody (1:400 dilution; EPR3894, GeneTex, San Antonio, TX, United States), and the rabbit anti-MSH2 polyclonal antibody (1:200 dilution; 15520-1-AP, Proteintec, Chicago, IL, United States). Samples were treated overnight with each primary antibody at 4 °C. Immunostaining was performed blindly by an investigator (Fukuda K) who was unaware of the clinical information using an N-Histofine Simple Stain MAX-PO kit (Nichirei).

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