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.
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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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