This gene encodes an essential enzyme of nucleotide metabolism. The encoded protein forms a ubiquitous, homotetrameric enzyme that hydrolyzes dUTP to dUMP and pyrophosphate. This reaction serves two cellular purposes: providing a precursor (dUMP) for the synthesis of thymine nucleotides needed for DNA replication, and limiting intracellular pools of dUTP. Elevated levels of dUTP lead to increased incorporation of uracil into DNA, which induces extensive excision repair mediated by uracil glycosylase. This repair process, resulting in the removal and reincorporation of dUTP, is self-defeating and leads to DNA fragmentation and cell death. Alternative splicing of this gene leads to different isoforms that localize to either the mitochondrion or nucleus. A related pseudogene is located on chromosome 19.

推荐稀释比 WB: 1/1000-1/5000; FC: 1/200-1/2000; ICC/IF: 1/100-1/1000

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Western blotting analysis using DUT antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with DUT antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using DUT antibody . DUT expression in wild type (WT) and DUT shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with DUT antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Immunocytochemical staining of HeLa cells using DUT antibody , Top panel: wild-type (WT); Bottom panal: DUT shRNA knockdown (KD). Nuclei were stained blue with DAPI;DUT was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.

Flow cytometric analysis of DUT expression in HeLa cells using DUT antibody . Green, isotype control; red, DUT.

Immunocytochemical staining of Hela cells with DUT antibody (Cat #61945, 1:1,000) . Nuclei were stained blue with DAPI; DUT 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.

Validation of DUT knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with DUT antibody and analyzed using BD flow cytometer.

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