Background

Thymidylate synthase catalyzes the methylation of deoxyuridylate to deoxythymidylate using, 10-methylenetetrahydrofolate (methylene-THF) as a cofactor. This function maintains the dTMP (thymidine-5-prime monophosphate) pool critical for DNA replication and repair. The enzyme has been of interest as a target for cancer chemotherapeutic agents. It is considered to be the primary site of action for 5-fluorouracil, 5-fluoro-2-prime-deoxyuridine, and some folate analogs. Expression of this gene and that of a naturally occurring antisense transcript, mitochondrial enolase superfamily member 1 (GeneID:55556), vary inversely when cell-growth progresses from late-log to plateau phase. Polymorphisms in this gene may be associated with etiology of neoplasia, including breast cancer, and response to chemotherapy.

Images

	Western blotting analysis using anti-thymidylate synthetase antibody (Cat#61673). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-thymidylate synthetase antibody (Cat#61673, 1:5,000) and HRP-conjugated goat anti rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
	Western blotting analysis using anti-thymidylate synthetase antibody (Cat#61673). Thymidylate synthetase expression in wild type (WT) and thymidylate synthetase (TYMS) knockdown (KD) HSHC cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with anti-thymidylate synthetase antibody (Cat#61673, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
	Flow cytometric analysis of Thymidylate synthetase expression in HepG2 cells using anti-Thymidylate synthetase antibody (Cat#61673, 1:2,000). Green, isotype control; red, Thymidylate synthetase.
	Immunocytochemical staining of HepG2 cells with anti-Thymidylate synthetase antibody (Cat#61673, 1:1,000). Nuclei were stained blue with DAPI; Thymidylate synthetase was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.

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