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Glutaminase activity determination kit and method based on micromethod

A technology for measuring glutaminase and activity, which is applied in the field of life science and can solve the problems of large interference, inability to obtain, and unstable measurement results.

Inactive Publication Date: 2018-12-21
SUZHOU COMIN BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, the Nestle method lacks the following disadvantages: 1. The preparation is cumbersome and requires five reagents, of which HgCl 2 It is highly toxic and cannot be obtained in general laboratories; 2. The method is greatly disturbed and has false positives; 3. The measurement results are unstable and poor in repeatability

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  • Glutaminase activity determination kit and method based on micromethod
  • Glutaminase activity determination kit and method based on micromethod
  • Glutaminase activity determination kit and method based on micromethod

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Embodiment Construction

[0095] The present invention will be described in detail below in conjunction with embodiments.

[0096] A kind of glutaminase activity assay kit based on micromethod, comprising following reagents:

[0097] Reagent 1, PH8.0 disodium hydrogen phosphate-citric acid buffer solution 60 mL×2 bottles; each bottle of disodium hydrogen phosphate-citric acid buffer solution is composed of 0.1795g Na 2 HPO 4 , 0.0347g citric acid and 60mL distilled water are mixed, and placed in a 60mL volumetric flask, stored at 4°C;

[0098] Reagent 2, 0.05% glutamine aqueous solution 40 mL × 1 bottle; the glutamine aqueous solution is prepared by mixing 0.02 g glutamine and 40 mL distilled water, and placed in a 40 mL volumetric flask, stored at 4 °C;

[0099] Reagent 3, 20% TCA aqueous solution 60 mL×1 bottle; the TCA aqueous solution is made by mixing 12g TCA and 60mL distilled water, and placed in a 60mL volumetric flask, stored at room temperature;

[0100] Reagent 4, ninhydrin powder x 1 bot...

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Abstract

The invention discloses a glutaminase activity determination kit and method based on micromethod. The kit provided by the invention comprises a first reagent: disodium hydrogen phosphate-citric acid buffer solution, a second reagent: glutamine aqueous solution, a third reagent: TCA aqueous solution, and a fourth reagent: ninhydrin powder. According to a determination principle provided by the invention, glutamine is catalyzed by GLS and is dissolved into L-glutamic acid and ammonia, and an increase rate of the glutamic acid is determined through utilization of a ninhydrin development method for the first time, thereby indirectly reflecting GLS enzymatic activity, so the kit and the method have originality. The employed reagent: ninhydrin is simple in preparation and is a common reagent ina laboratory. According to the determination method provided by the invention, through reasonable setting of comparison tubes, influence of impurities can be eliminated, and detection specifity is very high. The determination method provided by the invention is relatively good in repeatability. A variable coefficient (CV value) is within 2%. The determination method provided by the invention is applicable to various samples such as animals, plants, microorganisms and culture cells and is wide in application range.

Description

technical field [0001] The invention belongs to the field of life sciences, and in particular relates to a kit for measuring glutaminase activity based on a trace method and a method thereof. Background technique [0002] GLS (EC 3.5.1.1) is glutaminase, which can catalyze the hydrolysis of glutamine into L-glutamic acid and ammonia, and plays an important role in nitrogen metabolism, especially in regulating free ammonia content and urea metabolism Regulation. The currently existing method for measuring glutaminase activity is the Nessler method, which uses the Nessler reagent to measure the increase in ammonia to reflect the level of glutaminase activity. [0003] However, the Nestle method lacks the following disadvantages: 1. The preparation is cumbersome and requires five reagents, of which HgCl 2 It is highly toxic and cannot be obtained in general laboratories; 2. The method is greatly disturbed and there are false positives; 3. The measurement results are unstable ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/31G01N21/78
Inventor 姚金美赵林川
Owner SUZHOU COMIN BIOTECH
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