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US 2014/0273036 A1Published application
Luminescent detection of inorganic phosphate and coupled reactions
AssigneePromega Corporation
InventorsSubhanjan Mondal, Kuei-Hsuan Hsiao, Said Goueli
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Abstract
Luminescent detection of inorganic phosphate is carried out in an assay through the intermediary enzymatic production of ADP. ADP is converted to ATP which is used in a luminescent reaction. The assay can be used to monitor coupled enzyme reactions which use or generate inorganic phosphate and the modulation of such reactions.
Claims22
A method for detecting the presence or absence of or determining the amount of inorganic phosphate (Pi) in a solution, the method comprising:
- (a) contacting the solution with polynucleotide phosphorylase to convert the inorganic phosphate to ADP;
- (b) converting the ADP produced from the inorganic phosphate to ATP with an enzyme that catalyzes the conversion of ADP to ATP;
- and
- (c) detecting the ATP produced from the ADP using a luminescent reaction.
1 more element
A kit for detecting inorganic phosphate in a solution, the kit comprising:
- (i) a polynucleotide phosphorylase;
- (ii) poly(A)
- (iii) an enzyme that catalyzes the conversion of ADP to ATP;
- (iv) a luciferase enzyme;
- and
- (v) a substrate for the luciferase enzyme.
3 more elements
Dependencies are inferred from the claim text.