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Kit for detecting activity of MTHFR (5,10-methylene tetrahydrofolate reductase)

A methylenetetrahydrofolate and kit technology, which is applied in the field of kits for detecting enzyme activity and kits for detecting enzyme activity of methylenetetrahydrofolate reductase, can solve problems such as difficulties in kinetic parameters of natural substrates

Inactive Publication Date: 2017-02-22
北京中科卓明生物医学研究所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] It has been reported in the literature that the HPLC method is used to detect the normal physiological response of MTHFR, but this method is used to detect the extracts of pig liver tissue and cultured fibroblasts, and the basic substrate must first be synthesized from formaldehyde and tetrahydrofolate in the reaction system 5,10-Methylenetetrahydrofolate, followed by subsequent reactions, which makes it difficult to precisely determine the kinetic parameters of the natural substrate

Method used

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  • Kit for detecting activity of MTHFR (5,10-methylene tetrahydrofolate reductase)
  • Kit for detecting activity of MTHFR (5,10-methylene tetrahydrofolate reductase)
  • Kit for detecting activity of MTHFR (5,10-methylene tetrahydrofolate reductase)

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Embodiment 1, the detection kit of methylenetetrahydrofolate reductase enzymatic activity, comprises the following components:

[0057] Reagent 1:

[0058] Potassium phosphate buffer (pH6.8) 80mmol / L

[0059] Dithiothreitol 5mmol / L

[0060] Reagent 2:

[0061] 5,10-methylenetetrahydrofolate aqueous solution 0.5mmol / L

[0062] Reagent 3:

[0063] NADPH aqueous solution 2mmol / L

[0064] Reagent 4:

[0065] Perchloric acid aqueous solution 60ml / L

[0066] Ascorbic acid aqueous solution 60mmol / L

[0067] Reagent 5:

[0068] Potassium phosphate buffer (pH6.8) 60mmol / L

[0069] Ascorbic acid aqueous solution 30mmol / L

Embodiment 2

[0070] Embodiment 2, the detection kit of methylenetetrahydrofolate reductase enzymatic activity, comprises the following components:

[0071] Reagent 1:

[0072] Potassium phosphate buffer (pH7.0) 100mmol / L

[0073] Dithiothreitol 5mmol / L

[0074] Reagent 2:

[0075] 5,10-methylenetetrahydrofolate aqueous solution 0.4mmol / L

[0076] Reagent 3:

[0077] NADPH aqueous solution 1.5mmol / L

[0078] Reagent 4:

[0079] Perchloric acid aqueous solution 60ml / L

[0080] Ascorbic acid aqueous solution 60mmol / L

[0081] Reagent 5:

[0082] Potassium phosphate buffer (pH6.8) 60mmol / L

[0083] Ascorbic acid aqueous solution 30mmol / L

Embodiment 3

[0084] Embodiment 3, the detection kit of methylenetetrahydrofolate reductase enzymatic activity, comprises the following components:

[0085] Reagent 1:

[0086] Potassium phosphate buffer (pH7.2) 150mmol / L

[0087] Dithiothreitol 5mmol / L

[0088] Reagent 2:

[0089] 5,10-methylenetetrahydrofolate aqueous solution 0.15mmol / L

[0090] Reagent 3:

[0091] NADPH aqueous solution 1mmol / L

[0092] Reagent 4:

[0093] Perchloric acid aqueous solution 80ml / L

[0094] Ascorbic acid aqueous solution 60mmol / L

[0095] Reagent 5:

[0096] Potassium phosphate buffer (pH6.8) 60mmol / L

[0097] Ascorbic acid aqueous solution 30mmol / L

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Abstract

The invention discloses a kit for detecting activity of MTHFR (5,10-methylene tetrahydrofolate reductase), a method and an application of the kit. The kit comprises a reagent 1, a reagent 2, a reagent 3, a reagent 4 and a reagent 5 which are packaged independently, wherein the reagent 1 is a buffer system formed by a buffer solution 1 and an enzyme activity stabilizer 1, and the pH value of the reagent 1 ranges from 5 to 8; the reagent 2 is methylene tetrahydrofolate; the reagent 3 is NADPH; the reagent 4 is a stop buffer formed by a stop reagent and an enzyme activity stabilizer 2; the reagent 5 is a diluent formed by a buffer solution 2 and the enzyme activity stabilizer 2, and the pH value of the reagent 5 ranges from 5 to 8. The method can be applied to clinics and physical examination screening, meets the requirements for physical examination screening, diagnosis and treatment of MTHFR deficiency, folic acid utilization obstacle and folic acid deficiency, and is easy to popularize and use.

Description

technical field [0001] The invention relates to a kit for detecting enzyme activity, in particular to a kit, method and application for detecting the enzyme activity of methylenetetrahydrofolate reductase, belonging to the field of biomedical detection. Background technique [0002] Folic acid is a methyl donor in the synthesis of many important substances in the body, such as protein, nucleic acid, hemoglobin, etc. The lack of folic acid or the lack of folic acid metabolic enzymes will lead to abnormal cell cycle, abnormal methylation reaction of DNA and protein, DNA base Abnormal biochemical reactions, such as the failure to synthesize normally, lead to many major diseases. [0003] The metabolism of folic acid is catalyzed by a series of enzymes. Folic acid must be converted into an active form of 5-methyltetrahydrofolate to complete its normal function in the body. The key enzyme to realize the conversion of this active substance is 5,10 -Methylenetetrahydrofolate reduc...

Claims

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Application Information

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IPC IPC(8): C12Q1/26
CPCC12Q1/26
Inventor 王天泽郝淑静
Owner 北京中科卓明生物医学研究所有限公司