DTYMK (deoxythymidylate kinase (thymidylate kinase)), also known as CDC8, TMPK, TYMK or dTMP kinase, is a 212 amino acid protein that belongs to the thymidylate kinase family and is involved in pyrimidine metabolism. Specifically, DTYMK catalyzes the ATP-dependent conversion of dTMP (deoxythymidine monophosphate) to dTDP (deoxythymidine diphosphate), which then functions as one of the four nucleotides in DNA. Via its role in the catalytic creation of dTDP, DTYMK plays an important role in the pathway of DNA synthesis and is thought to be involved in cell cycle progression and cell growth. DTYMK expression levels peak during the S phase (synthesis phase) of the cell cycle, further supporting the role of DTYMK in DNA synthesis.
Background References
1. Zhou T. et. al. DTYMK promote hepatocellular carcinoma proliferation by regulating cell cycle. Cell Cycle. 2021 Sep
2. Guo Y. et. al. DTYMK Expression Predicts Prognosis and Chemotherapeutic Response and Correlates with Immune Infiltration in Hepatocellular Carcinoma. J Hepatocell Carcinoma. 2021 Aug
Western blot analysis of DTYMK on different lysates with Rabbit anti-DTYMK antibody (HA721116) at 1/1,000 dilution.
Lane 1: HepG2 cell lysate Lane 2: Huh7 cell lysate Lane 3: MCF7 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 24 kDa Observed band size: 25 kDa
Exposure time: 20 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 (HA721116) 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.
Immunocytochemistry analysis of NCI-H441 cells labeling DTYMK with Rabbit anti-DTYMK antibody (HA721116) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-DTYMK antibody (HA721116) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-DTYMK antibody (HA721116) at 1/100 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 (HA721116) at 1/100 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.
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