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US 2010/0075350 A1Published application
ADP detection based luminescent phosphotransferase or ATP hydrolase assay
AssigneePromega Corporation
InventorsHicham Zegzouti, Said A. Goueli
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Abstract
The invention provides compositions and methods to determine or detect the activity of enzymes, including phosphotransferases such as kinases (e.g., protein, lipid, and sugar kinases) and ATP hydrolases such as ATPases, e.g., HSP90, that employ ATP as a substrate and form ADP as a product by monitoring changes in ADP.
Claims14
A method for detecting the presence or absence of or determining the amount of adenosine diphosphate (ADP) in a solution, comprising:
- (a) reducing substantially all ATP in the solution to a non-ADP product using an enzyme that converts ATP to a non-ADP product;
- (b) converting ADP in the solution to ATP using an enzyme that is capable of converting ADP to ATP and a phosphate group donor;
- and
- (c) detecting the presence or absence of or determining the amount of the ATP produced in (a) using a bioluminescent reaction.
1 more element
A kit for detecting ADP in a solution comprising:
- (i) an enzyme that converts a ATP to a non-ADP product;
- (ii) an enzyme that is capable of converting the ADP to ATP;
- (iii) a phosphate group donor;
- (iv) a luciferase enzyme;
- and
- (v) a substrate for the luciferase enzyme.
3 more elements
Dependencies are inferred from the claim text.