Method of measuring pyrophosphate
First Claim
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1. A method of determining the concentration of pyrophosphate in a sample, which method comprises the following steps(1) a step comprising a first reaction for producing ATP by contacting pyrophosphate which may be present in the sample with pyruvate orthophosphate dikinase in the presence of at least AMP;
- (2) a step comprising a second reaction for producing pyrophosphate and NAD by contacting the ATP produced in the first reaction with nicotinamide-nucleotide adenylyltransferase;
(3) a step comprising a third reaction for reducing the NAD produced by the second reaction into NADH;
(4) a step comprising detecting the amount of NADH produced by the third reaction; and
(5) determining the concentration of pyrophosphate in the sample based on the amount of NADH produced in the above step.
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Abstract
Provided is a novel and useful method of measuring pyrophosphate in a sample. There may be provided a novel and useful method of measuring pyrophosphate in a sample using pyruvate orthophosphate dikinase, nicotinamide-nucleotide adenylyltransferase, and dehydrogenase.
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Citations
19 Claims
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1. A method of determining the concentration of pyrophosphate in a sample, which method comprises the following steps
(1) a step comprising a first reaction for producing ATP by contacting pyrophosphate which may be present in the sample with pyruvate orthophosphate dikinase in the presence of at least AMP; -
(2) a step comprising a second reaction for producing pyrophosphate and NAD by contacting the ATP produced in the first reaction with nicotinamide-nucleotide adenylyltransferase; (3) a step comprising a third reaction for reducing the NAD produced by the second reaction into NADH; (4) a step comprising detecting the amount of NADH produced by the third reaction; and (5) determining the concentration of pyrophosphate in the sample based on the amount of NADH produced in the above step. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification