Monoamine oxidase enzyme activity detection kit and use method thereof

A monoamine oxidase enzyme and activity detection technology, applied in the biological field, can solve the problems of strict reagent storage conditions, incomplete sample extraction, large human error, etc., and achieve the effect of wide detection time range, reasonable sample extraction process and short detection time.

Pending Publication Date: 2021-06-08
ELABSCI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most of the monoamine oxidase enzyme activity detection kits on the market use the kynurenamide method, but it has the following disadvantages: incomplete sample extraction is prone to occur during the sample extraction process, human errors are large, the detection time is long, and the reagent storage conditions are strict. , The amount of reagents is large and the cost of reagents is high
The detection method is relatively complicated to operate, and benzyl aldehyde needs phenylhydrazine to be catalyzed to generate aldehyde phenylhydrazone, and phenylhydrazine itself is poisonous

Method used

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  • Monoamine oxidase enzyme activity detection kit and use method thereof
  • Monoamine oxidase enzyme activity detection kit and use method thereof
  • Monoamine oxidase enzyme activity detection kit and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A monoamine oxidase detection kit comprising the following reagents:

[0029] The sample extraction solution is 20mmol / L sucrose solution, and the solvent is double distilled water;

[0030] The enzyme extraction solution is 300mmol / L sucrose solution, and the solvent is double distilled water;

[0031] The substrate buffer is a PIPES buffer, wherein the PIPES concentration is 50mmol / L, the solvent is water, and the pH of the buffer is 6.5;

[0032] The chromogenic substrate is 4-p-dimethylaminobenzylamine hydrochloride, which can exist in the form of powder in a specific kit, or can be directly configured with matrix buffer to form a chromogenic substrate; in this example , the concentration of the chromogenic substrate in the chromogenic solution is 8mmol / L

[0033] The detection kit needs to be stored at a lower temperature, such as 4°C.

Embodiment 2

[0035] A monoamine oxidase detection kit comprising the following reagents:

[0036] The sample extraction solution is 10mmol / L sucrose solution, and the solvent is double distilled water;

[0037] Enzyme extraction liquid is the sucrose solution of 500mmol / L, and solvent is double distilled water;

[0038] The substrate buffer is a PIPES buffer, wherein the PIPES concentration is 30mmol / L, the solvent is water, and the pH of the buffer is 7.2;

[0039] The chromogenic substrate is 4-p-dimethylaminobenzylamine hydrochloride, which can exist in the form of powder in a specific kit, or can be directly configured with matrix buffer to form a chromogenic substrate; in this example , the concentration of the chromogenic substrate in the chromogenic solution is 5mmol / L

[0040] The detection kit needs to be stored at a lower temperature, such as 4°C.

Embodiment 3

[0042] Use the monoamine oxidase detection kit to detect the enzyme activity in the sample, which specifically includes the following steps:

[0043] (1) Preparation of samples to be tested

[0044] The processing methods applicable to different samples are different, and the following methods are used in this embodiment for processing:

[0045] ① Animal tissue sample processing

[0046] Take 0.1-0.5g sample, add the sample extract at the ratio of 0.1g:0.5-0.9mL for homogenization, centrifuge at 800-1000×g for 8-10min at 4°C, discard the precipitate; put the supernatant at 4°C Centrifuge at 9000-10000×g for 20-30min, discard the supernatant; then add 0.9-1mL enzyme extract solution, mix well, centrifuge at 15000-18000×g for 35-40min at 4°C, discard the supernatant; finally add 0.9-1mL matrix buffer Liquid working solution, after mixing, it is the sample to be tested, and it is stored in an ice box until use.

[0047] ② Cell sample processing

[0048] a. Collection of cultu...

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Abstract

The invention discloses a monoamine oxidase enzyme activity detection kit and a use method thereof. The kit comprises an extract, a matrix buffer solution and a chromogenic substrate, wherein the chromogenic substrate is 4-p-dimethylamino benzylamine hydrochloride, and the matrix buffer solution is a PIPES buffer solution. The detection kit is suitable for samples such as animal tissues, serum (plasma), cells and the like, and is simple and convenient to operate, short in detection time, low in detection cost, environment-friendly and non-toxic.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a monoamine oxidase enzyme activity detection kit and a use method thereof. Background technique [0002] Monoamine oxidase (MAO, EC1.4.3.4) is a flavoprotein that exists on the outer membrane of mitochondria and contains a sulfhydryl group. Its main function in the body is to catalyze the metabolism of endogenous and exogenous monoamines. Under the action of MAO, monoamines are oxidized to produce deamination. MAO is widely distributed in the central nervous system and nerve ending tissues, and is mainly located on the outer membrane of the mitochondria of tissue cells such as the brain, liver and intestinal mucosa. MAO is considered as an aging marker enzyme. With the increase of age, the activity of MAO in the human body increases, and abnormal MAO activity is related to neurological disorders, such as MAO-B and Parkinson's syndrome and Alzheimer's disease It is clo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573
CPCG01N33/573G01N2333/90638
Inventor 冷毅斌王长乐欧雄宇王振平陈冬琴
Owner ELABSCI BIOTECH
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