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Kit and method for determining content of glucose 6-phosphate (G6P) based on micro method

A glucose phosphate, micro-method technology, applied in the field of life sciences, can solve the problems of easy decomposition and unstable glucose 6-phosphate, and achieve the effect of good repeatability, simple and easy sample pretreatment process, and high accuracy

Inactive Publication Date: 2019-04-12
SUZHOU COMIN BIOTECH
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Problems solved by technology

However, there are currently few reports in the literature on the detection of the content of glucose 6-phosphate. The phosphate radicals contained in glucose 6-phosphate make glucose 6-phosphate unstable and easy to decompose. Therefore, a simple, fast, An effective method to detect its content

Method used

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Embodiment Construction

[0060] The present invention will be described in detail below in conjunction with embodiments.

[0061] A kit for the determination of glucose-6-phosphate content based on the micromethod, comprising the following reagents:

[0062] Extract, liquid 100 mL×1 bottle, composed of 100 mL, pH=7.4 Tris-HCl solution, stored at 4°C;

[0063] Reagent 1, liquid 21mL×1 bottle, composed of 0.254mgTris, 4.42mgMgCl 2 , 21mL of water and 22.1μL of concentrated hydrochloric acid, stored at 4°C;

[0064]Reagent 2, powder x 1 bottle, composed of 0.6mg glucose 6-phosphate dehydrogenase (G6PDH), stored at -20°C;

[0065] Reagent 3, powder x 1 bottle, composed of 42mg NADP, stored at -20°C.

[0066] A method for determining the content of glucose-6-phosphate based on the micro-method using the above-mentioned kit. The determination principle is that glucose-6-phosphate dehydrogenase (G6PDH) can catalyze glucose-6-phosphate (G6P) and NADP+ into NADPH at 340 nm Determination of NADPH increase r...

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Abstract

The invention discloses a kit and method for determining content of glucose 6-phosphate (G6P) based on a micro method. The kit comprises the following reagents: an extract consisting of a Tris-HCl solution, a first reagent consisting of Tris, MgCl2, water and concentrated hydrochloric acid, a second reagent consisting of glucose-6-phosphate 1-dehydrogenase (G6PDH) and a third reagent consisting ofNADP. A principle of the method is as follows: the G6PDH can catalyze the G6P and NADP+ to NADPH and an increase rate of the NADPH at 340 nm is measured to reflect level of G6P content. The used micro method is rapid and effective in test and can prevent a problem of G6P decomposition possibly caused by too long test time. A sample pretreatment process is simple and easy, besides, measurement operation steps are all automatic operations of instruments, accurate data are read, each sample has a short test time, and the kit is good in repeatability and high in accuracy.

Description

technical field [0001] The invention belongs to the field of life sciences, and in particular relates to a kit and method for determining the content of glucose-6-phosphate based on a trace method. Background technique [0002] Glucose 6-phosphate (G6P) is an intermediate product of sugar metabolism and gluconeogenesis pathway, which widely exists in animals, plants and microorganisms. However, there are currently few reports in the literature on the detection of the content of glucose 6-phosphate. The phosphate radicals contained in glucose 6-phosphate make glucose 6-phosphate unstable and easy to decompose. Therefore, a simple, fast, An effective method is used to detect its content. Contents of the invention [0003] In order to meet the above needs, the present invention aims to provide a kit and method for determining the content of glucose-6-phosphate based on the micro-method, which has the characteristics of simple operation, fast and effective, good repeatability...

Claims

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Application Information

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IPC IPC(8): C12Q1/54C12Q1/32
CPCC12Q1/32C12Q1/54G01N2333/904
Inventor 焦岑蕾王跃赵林川
Owner SUZHOU COMIN BIOTECH
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