A kind of catechol-o-methyltransferase enzyme activity detection kit

A technology of methyltransferase and catechol, which is applied in the fields of biomedicine, clinical diagnosis and detection, and can solve the problems that the detection method of enzyme activity is not easy to popularize.

Active Publication Date: 2019-01-25
山西盈科正欣生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Therefore, based on the defects of the above-mentioned prior art, in order to solve the problem that the enzyme activity detection method of existing catechol-O-methyltransferase (COMT, Catechol-O-methyltransferase, EC 2.1.1.6) is not easy to popularize, the present invention The purpose is to provide a rapid, specific, accurate and reliable combination product for detecting COMT enzyme activity

Method used

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  • A kind of catechol-o-methyltransferase enzyme activity detection kit
  • A kind of catechol-o-methyltransferase enzyme activity detection kit
  • A kind of catechol-o-methyltransferase enzyme activity detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Embodiment 1: Preparation of human red blood cell lysate

[0087] Take 1ml of peripheral venous whole blood from healthy volunteers, centrifuge at 3000rpm for 10min, discard the supernatant, take 100ul of red blood cell pellet at the bottom, add 3ml of normal saline (0.9% NaCl aqueous solution) and shake well, centrifuge at 3000rpm for 10min, discard the supernatant, and then Wash twice with normal saline, centrifuge at 3000rpm for 10min, discard the supernatant, and the collected precipitate is red blood cells. The collected red blood cells can be stored at -40°C for future use. Add 2 volumes of 20mmol / L Tris-HCl buffer solution, pH 7.5, to one portion of red blood cell pellet, stir to break up the red blood cells, centrifuge at 10,000g for 10 minutes, and take the supernatant as red blood cell lysate.

Embodiment 2

[0088] Embodiment 2: Enzyme activity assay of catechol-O-methyltransferase

[0089] The catechol-O-methyltransferase detection reagent of this embodiment is three reagents, including

[0090] Reagent 1

[0091] Phosphate buffer (pH7.8) 30mmol / L

[0092] SAM0.2mmol / L

[0093] Reagent 2

[0094] Phosphate buffer (pH7.8) 30mmol / L

[0095] DBA0.4mmol / L

[0096] Reagent 3

[0097] Phosphate buffer (pH7.8) 30mmol / L

[0098] S-adenosyl-L-homocysteine ​​hydrolase 10KU / L

[0099] The sample of catechol-O-methyltransferase to be tested is the red blood cells of healthy adults. After crushing, sodium cholate solution is added to the sample to make the final volume percentage concentration reach 0.5%. After mixing gently, Place on ice for 30 min before use. The total protein dosage in the reaction system is 50ug, and the reaction system is 250ul. A total of 3 parallel samples were made. This detection method adopts the endpoint method to detect the enzyme activity of catechol...

Embodiment 3

[0106] Example 3: Determination of human red blood cell catechol-O-methyltransferase activity (comparison of pre-treatment of samples)

[0107] The catechol-O-methyltransferase detection reagent of this embodiment is three reagents, including

[0108] Reagent 1

[0109] Phosphate buffer (pH7.8) 30mmol / L

[0110] SAM0.2mmol / L

[0111] Reagent 2

[0112] Phosphate buffer (pH7.8) 30mmol / L

[0113] DBA0.4mmol / L

[0114] Reagent 3

[0115] Phosphate buffer (pH7.8) 30mmol / L

[0116] S-adenosyl-L-homocysteine ​​hydrolase 10KU / L

[0117] The sample of catechol-O-methyltransferase to be tested is mixed red blood cells of healthy adults. After reaching 0.5%, mix gently and place on ice for 30 minutes; add the same amount of sample suspension to the other part, mix gently and place on ice for 30 minutes. The total protein dosage in the reaction system is 25ug, and the reaction system is 250ul. This detection method adopts the endpoint method to detect the enzyme activity of ca...

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Abstract

The invention provides a combined product for detecting activity of catechol-O-methyl transferase. The combined product comprises S-adenosine-L-methionine or an enzyme reaction system generating S-adenosine-L-methionine, and the enzyme reaction system comprises triphosadenine, methionine, S-adenosine methionine synthetic enzyme, methyl receptor and S-adenosine-L-homocysteine hydrolase. Preferably, the combined product is in the form of a kit, and each ingredient is in a state of reagent existing independently. The combined product can be used on semi-automatic or full-automatic analysis and detection equipment, and is high in detection sensitivity, high in specificity and simple and convenient to operate, thereby being capable of being popularized and applied truly.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and at the same time belongs to the field of clinical diagnosis and detection. Specifically, the present invention provides a combination product for detecting catechol-O-methyltransferase (COMT, Catechol-O-methyltransferase, EC2.1.1.6) enzyme activity. Background technique [0002] Catechol-O-methyltransferase (COMT, Catechol-O-methyltransferase, EC 2.1.1.6), is an important metabolic enzyme widely present in mammalian tissues and red blood cells. -bound COMT, MB-COMT) and soluble (soluble COMT, S-COMT) two forms. The substrates of the methylation reaction of COMT in the human body are various catechol compounds, such as epinephrine, norepinephrine and dopamine (DA). In the central nervous system, COMT acts as a metabolic enzyme to inactivate neurotransmitters with a catechol structure; at the same time, it also metabolizes catechol estrogens, catechol drugs and some carcinogens. [0003]...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/48C12Q1/34
CPCC12Q1/34C12Q1/48G01N2333/46G01N2333/91017G01N2333/914G01N2800/2835G01N2800/30G01N2800/302G01N2800/304
Inventor 菅晓勇王天泽
Owner 山西盈科正欣生物科技有限公司
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