APPLICATION
Application Note
Thymidylate synthetase (TYMS) is an enzyme that catalyzes the conversion of deoxyuridine monophosphate (dumP) to deoxythymidine monophosphate (dTMP). 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. Thymidylate synthase is an enzyme of about 30 to 35kDa in most species except in protozoan and plants where it exists as a bifunctional enzyme that includes a dihydrofolate reductase domain. Besides, Aspartate Aminotransferase (AST) has been identified as an interactor of TYMS, thus a binding ELISA assay was conducted to detect the interaction of recombinant human TYMS and recombinant human AST. Briefly, TYMS were diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to AST-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-TYMS pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of TYMS and AST was in a dose dependent manner.
Observed MW
36 kDa.
PROPERTIES
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris (pH8.0) and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris (pH8.0), 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Gln36~Val313 (NP_001062.1)
Expression System
E. coli
Purity
> 90%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
thymidylate synthetase , HST422 , TMS , TS
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. [provided by RefSeq, Aug 2017]
Database
Research Area
DATA IMAGES
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GTX00420-pro Functional Assay Image
Functional ELISA analysis of GTX00420-pro Human Thymidylate synthase protein which can bind immobilized AST protein.
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GTX00420-pro Image
SDS-PAGE analysis of GTX00420-pro Human Thymidylate synthase protein.
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GTX00420-pro Image
WB analysis of GTX00420-pro Human Thymidylate synthase protein.
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REFERENCE
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REVIEW
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