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Determination method for S-adenosylmethionine methyltransferase and kit thereof

A technology for the determination of adenosylmethionine methyl group, which is applied in the field of analysis and detection technology of S-adenosylmethionine (SAM) methyltransferase (S-adenosylmethionine), It can solve the problems of time-consuming and laborious operation, cumbersome operation, pollution of the environment and operators, etc., and achieve the effect of convenient use, overcoming cumbersome operation and eliminating product accumulation.

Active Publication Date: 2012-10-03
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above-mentioned deficiencies of the prior art, the present invention proposes a method that removes the feedback inhibition of the product, has stable and reliable results, and accurate quantitative response, and overcomes the cumbersome operation, time-consuming and labor-intensive operation of the radioactive labeling method, and the radioactive effect on the environment and operators. Method for the detection of contaminating deficient SAM methyltransferase

Method used

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  • Determination method for S-adenosylmethionine methyltransferase and kit thereof
  • Determination method for S-adenosylmethionine methyltransferase and kit thereof
  • Determination method for S-adenosylmethionine methyltransferase and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Phosphate buffer for buffer, BSA for stabilizer

[0058] Reagent 1 is: 100mmol / L phosphate buffer, pH7.4

[0059] 20mmol / L MgCl 2

[0060] 50mmol / L S-adenosylmethionine

[0061] 50mmol / L adenosine triphosphate

[0062] 10mmol / L L-homocysteine

[0063] 1mmol / L TOOS

[0064] 1g / L BSA

[0065] 1g / L sodium azide

[0066] 10ml / L Triton X-100

[0067] Reagent 2 is: 100mmol / L phosphate buffer

[0068] 10mmol / L 4-aminoantipyrine

[0069] 100U / L S-adenosyl homocysteine ​​hydrolase

[0070] 100U / L Adenosine deaminase

[0071] 50U / L Purine nucleoside phosphorylase

[0072] 50U / L xanthine oxidase

[0073] 100U / L Peroxidase

[0074] 1g / L BSA

[0075] 1g / L sodium azide

[0076] 10ml / L Triton X-100;

[0077] The above kit is used for the enzyme coupling analysis and detection technology of SAM methyltransferase. The specific process is: through coupling with S-adenosyl homocysteine ​​hydrolase and adenosine deaminase, the SAM methyltransferase The generated product S...

Embodiment 2

[0080] Tris buffer for buffer, EDTA-Na for stabilizer 2

[0081] Reagent 1 is: 100mmol / LTris buffer, pH7.4

[0082] 20mmol / L MgCl 2

[0083] 50mmol / L S-adenosylmethionine

[0084] 50mmol / L adenosine triphosphate

[0085] 10mmol / L L-homocysteine

[0086] 1mmol / L TOOS

[0087] 1g / L sodium edetate

[0088] 1g / L sodium azide

[0089] 10ml / L Triton X-100

[0090] Reagent 2 is: 100mmol / LTris buffer

[0091] 10mmol / L 4-aminoantipyrine

[0092] 100U / L S-adenosyl homocysteine ​​hydrolase

[0093] 100U / L Adenosine deaminase

[0094] 50U / L purine nucleoside phosphorylase

[0095] 50U / L xanthine oxidase

[0096] 100U / L peroxidase

[0097] 1g / L sodium edetate

[0098] 1g / L sodium azide

[0099] 10ml / L Triton X-100

[0100] The assay method of embodiment 2 is identical with embodiment 1.

Embodiment 3

[0102] Glycine buffer was used as the buffer, and mannitol was used as the stabilizer.

[0103] Reagent 1 is: 100mmol / L glycine buffer solution, pH7.4

[0104] 20mmol / L MgCl 2

[0105] 50mmol / L S-adenosylmethionine

[0106] 50mmol / L adenosine triphosphate

[0107] 10mmol / L L-homocysteine

[0108] 1mmol / L TOOS

[0109] 1g / L Mannitol

[0110] 1g / L sodium azide

[0111] 10ml / L Triton X-100

[0112] Reagent 2 is: 100mmol / L glycine buffer

[0113] 10mmol / L 4-aminoantipyrine

[0114] 100U / L S-adenosyl homocysteine ​​hydrolase

[0115] 100U / L Adenosine deaminase

[0116] 50U / L purine nucleoside phosphorylase

[0117] 50U / L xanthine oxidase

[0118] 100U / L peroxidase

[0119] 1g / L Mannitol

[0120] 1g / L sodium azide

[0121] 10ml / L Triton X-100

[0122] The assay method of embodiment 3 is identical with embodiment 1.

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Abstract

The invention discloses a determination method for S-adenosylmethionine methyltransferase and a kit thereof. The determination method for S-adenosylmethionine methyltransferase comprises the step of detecting SAM methyltransferase through the enzyme coupling reaction between S-adenosylhomocysteine hydrolase and adenosine deaminase. The detection technique relieves feedback inhibition of products,has stable results and accurate quantitative reaction, and effectively overcomes the problem of radioactive pollution of the radioactive marking method. With double reagents in the kit, detection canbe carried out manually or through a full automatic biochemical analyzer within the visible range, and therefore the use and the fast detection of a plurality of samples are convenient. The determination method is also suitable for detection of the enzyme coupling reaction between S-adenosylhomocysteine hydrolase and adenosine deaminase.

Description

Technical field: [0001] The invention relates to the fields of biochemistry and test reagents, in particular to an enzyme couple of S-adenosylmethionine (SAM) methyltransferase (S-adenosylmethionine-dependent methyltransferase) Combined analysis and detection technology, and the detection kit developed based on this technology, the kit can also be prepared into S-adenosyl homocysteine ​​hydrolase or adenosine deaminase detection kit after improvement. Background technique: [0002] The methylation modification of biological macromolecules plays an important role in signal transduction, biosynthesis, protein repair, gene silencing and regulation of chromosome expression. S-adenosyl-L-methionine (SAM) It is the methyl donor second only to ATP in the body, and its dependent S-adenosylmethionine transferase uses SAM as a methyl donor to methylate DNA and proteins, thereby participating in important physiological activities . Current studies have shown that the abnormal level o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48C12Q1/34C12Q1/28C12Q1/26
Inventor 邹炳德邹继华贾江花章玉胜
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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