Thymidylate synthase (TS) (EC 2.1.1.45) is an enzyme that catalyzes the conversion of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP). Thymidine is one of the nucleotides in DNA. With inhibition of TS, an imbalance of deoxynucleotides and increased levels of dUMP arise. Both cause DNA damage. By means of reductive methylation, deoxyuridine monophosphate (dUMP) and N5,N10-methylene tetrahydrofolate are together used to form dTMP, yielding dihydrofolate as a secondary product. This provides the sole de novo pathway for production of dTMP and is the only enzyme in folate metabolism in which the 5,10-methylenetetrahydrofolate is oxidised during one-carbon transfer. The enzyme is essential for regulating the balanced supply of the 4 DNA precursors in normal DNA replication: defects in the enzyme activity affecting the regulation process cause various biological and genetic abnormalities, such as thymineless death.
Background References
1. Monica V et al. Dasatinib modulates sensitivity to pemetrexed in malignant pleural mesothelioma cell lines. Oncotarget 7:76577-76589 (2016).
2. Lund K et al. Slug-dependent upregulation of L1CAM is responsible for the increased invasion potential of pancreatic cancer cells following long-term 5-FU treatment. PLoS One 10:e0123684 (2015).
Western blot analysis of Thymidylate Synthase on different lysates with Rabbit anti-Thymidylate Synthase antibody (ET1705-24) at 1/1,000 dilution.
Lane 1: NIH/3T3 cell lysate (20 µg/Lane) Lane 2: RAW264.7 cell lysate (20 µg/Lane) Lane 3: C6 cell lysate (20 µg/Lane) Lane 4: HeLa cell lysate (20 µg/Lane) Lane 5: Mouse thymus tissue lysate (40 µg/Lane)
Predicted band size: 36 kDa Observed band size: 35 kDa
Exposure time: Lane 1-3: 3 minutes; Lane 4-5: 46 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-24) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Thymidylate Synthase antibody (ET1705-24) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-24) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-Thymidylate Synthase antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded rat bone tissue using anti-Thymidylate Synthase antibody. Counter stained with hematoxylin.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"