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Luciferase freeze-dried powder dilution reaction liquid

A technology of luciferase and reaction solution, applied in the directions of enzyme stabilization, oxidoreductase, etc., can solve the problems of limited application prospects of luciferase, and achieve the effect of simple formula

Pending Publication Date: 2022-04-15
合肥巅峰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the principle of luciferase catalyzed luminescence is basically known by researchers, the instability of luciferase limits its application prospects, so the main research work of most researchers is on the study of stability. , such as the study of protective agents or the expression of more stable luciferase by site-directed mutagenesis

Method used

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  • Luciferase freeze-dried powder dilution reaction liquid
  • Luciferase freeze-dried powder dilution reaction liquid
  • Luciferase freeze-dried powder dilution reaction liquid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A kind of luciferase freeze-dried powder dilution reaction solution is provided in embodiment 1, comprises following components: luciferin 1mM, PEG1mM, Tris 0.1mM, HEPPS 1mM, magnesium chloride 0.25mM, potassium chloride 0.01mM, sodium chloride 0.05mM .

[0020] Among them, the detection method includes: taking 100MG of luciferase freeze-dried powder in a centrifuge tube, dissolving and diluting to 10ML with the prepared dilution buffer, taking 10ul 1×10 -9 Put the M ATP standard solution in the reaction tube, add 500ul of the dissolved reaction solution, put it into the handheld ATP bioluminescence detector for detection, and record the detection data. Repeat the above detection steps 10 times to obtain 10 sets of data. The results are summarized in Table 1 below:

[0021] Table 1: The detection result that draws in embodiment 1

[0022]

Embodiment 2

[0024] A kind of luciferase freeze-dried powder dilution reaction solution is provided in embodiment 2, comprises following components: luciferin 2mM, PEG0.5mM, Tris 0.1mM, HEPPS 1mM, magnesium chloride 0.25mM, potassium chloride 0.01mM, sodium chloride 0.05mM, Glycine 2.5mM. The test results are summarized in Table 2 below:

[0025] Table 2: The detection result that draws in embodiment 2

[0026]

Embodiment 3

[0028] A kind of luciferase freeze-dried powder dilution reaction solution is provided in embodiment 3, comprises following components: luciferin 2mM, PEG0.5mM, Tris 0.1mM, HEPPS 1mM, magnesium chloride 0.25mM, potassium chloride 0.01mM, sodium chloride 0.05mM, Glycine 2.5mM, Glutathione 10mM. The test results are summarized in Table 3 below:

[0029] Table 3: The detection result that draws in embodiment 3

[0030]

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Abstract

The invention provides a luciferase freeze-dried powder dilution reaction solution. The luciferase freeze-dried powder dilution reaction solution is prepared from the following components: 0.1 to 3 mM of fluorescein, 0.1 to 1 mM of polyethylene glycol, 0.1 to 1 mM of trihydroxymethyl aminomethane, 0.5 to 2 mM of 4-hydroxyethyl piperazine propanesulfonic acid, 0.1 to 0.5 mM of magnesium chloride, 0.01 to 0.3 mM of potassium chloride, 0.05 to 0.15 mM of sodium chloride, 0 to 3 mM of glycine and 0 to 20 mM of glutathione. The luciferase freeze-dried powder provided by the invention is used for diluting the reaction liquid, so that the stability of the luciferase dry powder is maintained, and the activity of the luciferase can be effectively reflected.

Description

technical field [0001] The invention relates to the technical field of luciferase dry powder, in particular to a diluted reaction solution of luciferase freeze-dried powder. Background technique [0002] Firefly luciferase catalyzes luminescence in the presence of ATP, luciferin and Mg 2+ In the presence of yellow-green fluorescence. Generally, most scientists believe that luciferase is used as a catalyst to catalyze D-luciferin, and D-luciferin oxidizes and emits light, and under certain conditions, the emission intensity of fluorescence is directly related to the concentration of ATP. [0003] Although the principle of luciferase catalyzed luminescence is basically known by researchers, the instability of luciferase limits its application prospects, so the main research work of most researchers is on the study of stability. , such as the study of protective agents or the expression of more stable luciferase through site-directed mutagenesis. Luciferase dry powder has go...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N9/96
Inventor 马振桥丁海平李坤
Owner 合肥巅峰生物科技有限公司