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Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is colon carcinoma.
Thymidylate Synthase (TS) converts deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP), which is essential for DNA biosynthesis. TS is a critical target for the fluoropyrimidines, an important group of antineoplastic drugs that are widely used in the treatment of solid tumors. Both 5-FU and fluorodeoxyuridine are converted in tumor cells to FdUMP which inactivates TS by formation of a ternary covalent complex in the presence of the folate cofactor 5,10-methylenetetrahydrofolate. Expression of TS protein is associated with response to 5-fluorouracil (5-FU) in human colorectal, gastric, head and neck, and breast carcinomas.
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