Publication protocol
"CDH17 mouse monoclonal (1:4000; clone IH3, Biocare Medical, Concord, California), CK20 (1:10 000; Ks20.8, Biocare Medical), and CDX2 (1:250, CDX2-88; BioGenex, Fremont, California) antibodies were evaluated with immunohistochemistry for specificity and sensitivity on formalin-fixed, paraffin-embedded tissue microarrays (TMAs) consisting of normal tissues (n = 26), colon adenocarcinoma (n = 149), stomach adenocarcinoma (n = 175), esophageal cancer (n = 54), appendix cancer (n = 5), pancreatic cancer (n = 57), hepatocellular carcinoma (n = 57), cholangiocarcinoma (n = 12), ovarian cancer (n = 60), endometrial adenocarcinoma (n = 7), lung cancer (n = 78), prostate adenocarcinoma (n = 20), breast cancer (n = 73), bladder cancer (n = 63), renal cell carcinoma (n = 10), thyroid cancer (n = 12), seminoma (n = 23), brain cancer (n = 12), melanoma (n = 6), and lymphoma (n = 11). Antibody titers were optimized and applied for 30 minutes on TMA specimens, followed by a polymer-based horseradish peroxidase detection system (MACH 4 HRP-Polymer, Biocare Medical). Staining was visualized with 3,3′-diaminobenzidine chromogen (Betazoid DAB Chromogen Kit, Biocare Medical).
To compare positive staining of CDH17 in tissues with CK20 and CDX2, statistical analyses were performed with GraphPad Prism version 6.0 (GraphPad Software, La Jolla, California). P values were determined by using the χ2 test, and values less than or equal to 0.05 were considered statistically significant."
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