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A glutathione reductase assay kit and its preparation method and application

A technology of glutathione and reductase, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor anti-interference ability, inconvenient use, reconstitution, etc., and achieve the effect of improving accuracy

Active Publication Date: 2019-05-14
中拓生物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the methods for the determination of glutathione reductase in China mainly include Elisa method and ultraviolet enzymatic method. The former requires manual operation, takes a long time, and the steps are cumbersome and expensive.
At present, the method mainly promoted in China is the ultraviolet enzymatic method, but the existing kits for ultraviolet enzymatic glutathione reductase are mainly imported dry powder reagents. Although the results are accurate, they are expensive and require reconstitution before use, which is difficult to operate. convenient
It needs to be used as soon as possible after reconstitution, it is very inconvenient to use and there will be a lot of waste if it is not used up after reconstitution
However, the domestic liquid dual reagents that are most convenient to use on clinical biochemical analyzers currently have disadvantages such as poor stability and poor anti-interference ability.

Method used

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  • A glutathione reductase assay kit and its preparation method and application
  • A glutathione reductase assay kit and its preparation method and application
  • A glutathione reductase assay kit and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The components of reagent R1 are: Tris buffer 100mmol / L, EDTA 0.5mmol / L, pyruvate carboxylase 1.5KU / L, ascorbate oxidase 2KU / L, potassium ferrocyanide 6mg / L, Tween 20 1ml / L, sodium azide 0.5g / L;

[0042] The components in reagent R2 are: Tris buffer 100mmol / L, EDTA 0.5mmol / L, GSSG 2mmol / L, NADPH0.2mmol / L, soluble starch 1g / L, trehalose 1g / L, glycerin 4g / L, Sodium nitrogen 0.5g / L.

[0043] The pH value of reagent R1 is 7.0; the pH value of reagent R2 is 9.5;

[0044] Preparation method: (1) Preparation of reagent R1: take an appropriate amount of water, add the raw materials shown in R1 respectively, stir well to dissolve one raw material, then add the next raw material, adjust to the required pH value with hydrochloric acid or sodium hydroxide, and constant volume to required volume. (2) Preparation of reagent R2: Take an appropriate amount of water, add the raw materials shown in R1 respectively, stir well to dissolve one raw material, then add the next raw materia...

Embodiment 2

[0046]The components of reagent R1 are: Tris buffer 80mmol / L, EDTA 0.8mmol / L, pyruvate carboxylase 1.2KU / L, ascorbate oxidase 2.5KU / L, potassium ferrocyanide 8mg / L, brij35 1ml / L, sodium azide 0.5g / L;

[0047] The components in reagent R2 are: Tris buffer 80mmol / L, EDTA 0.8mmol / L, GSSG 2mmol / L, NADPH0.2mmol / L, soluble starch 1.5g / L, trehalose 1.5g / L, glycerol 6g / L , Sodium azide 0.5g / L.

[0048] The pH value of reagent R1 is 6.5; the pH value of reagent R2 is 9.3;

[0049] The preparation method of embodiment 2 kit is the same as embodiment 1

Embodiment 3

[0051] The components of reagent R1 are: Tris buffer 150mmol / L, EDTA 1mmol / L, pyruvate carboxylase 2KU / L, ascorbate oxidase 3KU / L, potassium ferrocyanide 10mg / L, TritonX-100 1ml / L , sodium azide 0.5g / L;

[0052] The components in reagent R2 are: Tris buffer 150mmol / L, EDTA 1mmol / L, GSSG 2mmol / L, NADPH0.2mmol / L, soluble starch 2g / L, trehalose 2g / L, glycerol 9g / L, azide Sodium 0.5g / L.

[0053] The pH value of reagent R1 is 7.0; the pH value of reagent R2 is 9.0;

[0054] The preparation method of embodiment 3 kit is the same as embodiment 1

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Abstract

The invention provides a glutathione reductase assay kit, which comprises a reagent R1 and a reagent R2. The reagent R1 includes: 50-200 mmol / L of a Tris buffer solution, 0.5-2 mmol / L of EDTA, 1-5 kU / L of pyruvic carboxylase, 1-5 kU / L of ascorbic acid oxidase, 5-20 mg / L of potassium ferrocyanide, a surfactant and a preservative; the reagent R2 includes: 50-200 mmol / L of the Tris buffer solution, 0.5-2 mmol / L of EDTA, 2-10 mmol / L of GSSG, 0.1-0.5 mmol / L of NADPH, 1-15 g / L of a stabilizer, and 0.5-2 g / L of a preservative. The liquid assay kit has good stability and is strong in anti-interferenceeffect. The invention also discloses a preparation method and an application of the assay kit.

Description

technical field [0001] The invention relates to the technical field of biochemical reagent assay, in particular to a glutathione reductase assay kit, and also to a preparation method and application of the glutathione reductase assay kit. Background technique [0002] Glutathione reductase (GR) is a flavin enzyme, and each molecule of enzyme protein contains one molecule of FAD. The coenzyme NADPH supplies hydrogen to catalyze the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH). Reduced glutathione (GSH) can make enzymes containing sulfhydryl groups (-SH) in reduced and active states, maintain the integrity of red blood cell membranes, and prevent hemoglobin oxidation. The activity of GR in the body is strong and weak, followed by liver, kidney, pancreas, heart, thyroid, red blood cells and plasma. GR localized in microsomes and cytosol. Because the tissue cells of various organs generally contain GR, GR plays a pivotal role in the redox reaction of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573
CPCG01N33/573
Inventor 隗勇刘安娜张强
Owner 中拓生物有限公司
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