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Enzyme Activity Measurement of DNA Nucleotidylexotransferase

Creative Enzymes enjoys a good reputation in the enzyme testing industry. We have focused on transferases testing for years. Based on the standardized protocols and skillful operations, the testing results are of high accuracy and reproducibility. Creative Enzymes is the only company that provides rapid and precise activity assays for DNA nucleotidylexotransferase.

DNA nucleotidylexotransferase (TDT, EC 2.7.7.31) was first isolated from calf thymus in 1960. It is unique since it is the only known DNA polymerase that elongates DNA strands in a template-independent manner. In cell-free solutions, it catalyzes oligodeoxynucleotide (primer) extension by random selection of 3’-deoxyribonucleoside 5’-triphosphates (dNTPs) present in the incubation medium. Mg2+ is essential for the catalytic activity. TDT can achieve in vitro combination for not only ribonucleotides and deoxyribonucleotides but also a large number of unnatural nucleoside triphosphates.

Up till now, many reports have been published on TDT for its structures, functions, and genetic analyses. It is believed that the appearance of TDT seems to coincide with the emergence of an adaptive immune system. In normal animals, its strict limitation to primitive lymphoid cells suggests a role for generating the functional properties of T-cells and B-cells. Hence, a detailed investigation of the molecular mechanism of TDT action, as well as of inhibitors of this enzyme, are of interest both for basic molecular biology and for potential clinical applications. Recently, some applications are developed for this enzyme on analysis, biotechnology, and medical research. For example, TDT is used in staining of apoptotic cells of leukemia cell lines. With the increasing number of studies targeted at TDT, there is a notably growing number of inquiries on enzyme activity testing. Due to the unsteadiness in its properties and functions, quantification for this enzyme needs further investigation. Creative Enzymes has powerful platforms which can provide suitable activity testing according to various conditions.

Figure: The crystal structure of TDT in  complex with a DSB substrate and C-G base pair from mouse. Figure: The crystal structure of TDT in complex with a DSB substrate and C-G base pair from mouse.
Reference: Gouge, J. et al. Embo J. 2015 34: 1126-1142.

Creative Enzymes is a pioneer for DNA nucleotidylexotransferase activity testing. The comprehensive knowledge and efficient operations will guarantee precise results. Creative Enzymes will always keep a professional manner to every assay.



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