Product Overview :
ADK Phosphorylation Assay Kit was specially designed to evaluate the phosphorylation of novel nucleoside analogues. The assay
allows the test of 12 analogues at 7 different concentrations at the same time (or 6 analogues in duplicata).
This assay is based on competitive inhibition of inosine (IR) phosphorylation by ADK enzyme. In the absence of nucleoside competitor, adenosine kinase phosphorylates inosine resulting in the formation of IMP. IMP is further oxidized by IMPDH to XMP and NADH2.This coupling reaction is immediate when IMPDH activity is much higher than ADK activity in the assay. The enzymatic activity of ADK, which corresponds to the formation kinetics of IMP, is then stoichiometrically and directly monitored by the formation kinetics of NADH2. In the presence of nucleoside competitor, the phosphorylation of inosine, poor ADK substrate, is inhibited detected as a decrease in NADH2 formation.
This assay was validated with antiviral nucleoside analogues such as mizoribine, tubercidine, ribavirin.
Description :
Human adenosine kinase enzyme is a purine salvage enzyme that has a broad substrate specificity and provides the phosphorylation of purine/pyrimidine nucleosides and pyrimidine ribonucleosides. Adenosine kinase is critical for phosphorylation of adenosine analogs and provides first steps of activation of highly effective anti-viral and immunosuppressive prodrugs, such as mizoribine, tubercidin and ribavirine.
Storage :
ADK Phosphorylation Assay Kit must be stored at -20°C until used.
Size :
1 plate (96 assays)
Kit Components :
A standard ADK Phosphorylation Assay Kit (one 96-well plate) contains:
1. one tube "IMPDH";
2. one tube "Cofactor 1";
3. one tube "Cofactor 2";
4. one tube "Cofactor 3";
5. one vial "Reaction Buffer 5x";
6. one tube "Human ADK enzyme";
7. Transparent 96-well plate.
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