The Nucleoside Kinase Kit + Phosphate Regeneration System was designed to enzymatically phosphorylate a wide range of purine and pyrimidine nucleoside analogs using enzyme NK-12. The use of the phosphate donor regeneration system offers several advantages:
a) Simplified analytics and purification. Regeneration of the phosphate donor will - in case of 100 % conversion - lead to a two-compound reaction solution only containing the synthesized phosphorylated product of interest and ATP, whereas without regeneration also ADP and AMP will be present.
b) Free choice of substrate to phosphate donor ratio. Some enzymatic phosphorylation reactions require a low phosphate donor concentration. Ratios of substrate to phosphate donor lower than 1:1 will lead to a loss of yield if the phosphate donor is not regenerated during the reaction.
Biochemical action of nucleoside kinases:
Nucleoside kinases catalyze the following reaction:
Nucleoside + ATP ↔ NMP + ADP
Note: ATP is the preferred phosphate donor. However, if desired, other phosphate donors can be used, e.g. CTP. The included regeneration system works with all natural nucleoside triphosphates.
Unit definition: One unit of nucleoside kinase will cause the conversion of 1.0 μmol of substrate per min at pH 7.6 at optimum 37°C. ATP is the phosphate donor and is converted to ADP during the reaction. One unit of regeneration kinase will cause the conversion of 1.0 μmol of ADP to ATP per min at pH 7.6 at 37°C.
Product use limitations: This product is developed and sold exclusively for research purposes and in vitro use only. The product was not tested for use in diagnostics or drug development, nor is suitable for administration to humans or animals.