Kit and method for detecting activity of sucrose phosphate synthase

A phosphate synthase and activity determination technology, applied in the field of life sciences, can solve the problems of cumbersome operation process, inaccurate measurement results, large amount of reagents added, etc., achieve accurate measurement results, reduce the types of reagents, and reduce the cost of reagents

Inactive Publication Date: 2016-04-20
SUZHOU COMIN BIOTECH
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  • Abstract
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Problems solved by technology

[0004] 1. It is not suitable for the detection of samples with high sucrose content. The sucrose contained in the sample will have a great impact on the determination of the enzyme activity, resulting in inaccurate determination results
[0005] 2. The total reaction volume is 2-3mL, and the amount of various reagents added is relatively large, especially the concentration of UDPG is above 10mM. The reagent is expensive and the overall cost is high
[0006] 3. There are too many types of reagents and the operation process is too cumbersome

Method used

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  • Kit and method for detecting activity of sucrose phosphate synthase

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Embodiment Construction

[0062] The present invention will be described in detail below in conjunction with embodiments.

[0063] A sucrose phosphate synthase activity assay kit, comprising the following reagents:

[0064] Extract, liquid × 1 bottle, stored at 4°C, prepared by mixing 100mL of Tris-HCl buffer solution with a substance concentration of 100mM, pH=7.0, 203mg magnesium chloride, 58mgEDTA and 140uL mercaptoethanol, and placed in 100mL in the reagent bottle;

[0065] Reagent 1, liquid × 1 bottle, stored at -20°C, prepared by mixing 5mL of Tris-HCl buffer solution with a substance concentration of 50mM, pH=7.0, 10mg of magnesium chloride, 15.2mg of fructose-6-phosphate and 9mg of UDPG, and placed in In a 5mL reagent bottle;

[0066] Reagent 2, 1mg / mL sucrose solution 10mL×1 bottle, stored at 4°C, prepared by dissolving 10mg sucrose in 10mL distilled water, placed in a 10mL reagent bottle;

[0067] Reagent 3, liquid × 1 bottle, stored at 4°C, made by dissolving 0.16g of sodium hydroxide in ...

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Abstract

The invention discloses a kit and method for detecting the activity of sucrose phosphate synthase. The kit comprises an extracting solution prepared from Tris-HCl, magnesium chloride, EDTA and 2-mercaptoethanol, a reagent I prepared from Tris-HCl, magnesium chloride, 6-phosphofructose and UDPG, a reagent II prepared by dissolving saccharose in distilled water, a reagent III prepared by dissolving sodium hydroxide in distilled water, a reagent IV prepared by dissolving hydrochloric acid in distilled water and a reagent V prepared by dissolving resorcinol in ethanol and distilled water. According to the method, the influence of saccharose in a sample can be eliminated, so that the detection result is accurate, and the kit is suitable for detecting the activity of sucrose phosphate synthase in various samples. According to the kit and method, the number of reagent types can be greatly reduced, the operation can be simple and convenient, the total volume of the reaction is only 300mu L, the concentration of UDPG can be reduced on the base of not influencing the accuracy, and the reagent cost can be greatly reduced.

Description

technical field [0001] The invention belongs to the field of life sciences and relates to a kit, in particular to a kit for measuring the activity of sucrose phosphate synthase and a method thereof. Background technique [0002] Sucrose is not only an important photosynthetic product, but also the main substance transported in plants and one of the storage forms of carbohydrates. Sucrose phosphate synthase (SPS) accepts fructose-6-phosphate, and the formed sucrose phosphate is converted into sucrose under the action of sucrose phosphatase. Generally, the sucrose phosphate synthase-sucrose phosphatase system is regarded as the main pathway of sucrose synthesis. [0003] The currently available method for detecting the activity of sucrose phosphate synthase is the resorcinol colorimetric method. The principle is that sucrose phosphate synthase catalyzes fructose-6-phosphate to form sucrose phosphate in the sample, and the depth of the color change presented by the reaction b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
CPCC12Q1/48G01N2333/91125
Inventor 赵林川
Owner SUZHOU COMIN BIOTECH
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