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Adenosine triphosphate (ATP) fluorescence detection reagent

A fluorescence detection and reagent technology, applied in the field of ATP fluorescence detection, can solve the problems of poor thermal stability and short storage time, and achieve the effects of improving thermal stability, prolonging storage time, and preventing thermodynamic interference

Pending Publication Date: 2019-12-24
浙江东鸿生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the above-mentioned technical problems, provide a kind of ATP fluorescent detection reagent with good thermal stability, solve the problem that existing commercially available ATP fluorescent detection reagent has poor thermal stability and short storage time

Method used

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  • Adenosine triphosphate (ATP) fluorescence detection reagent
  • Adenosine triphosphate (ATP) fluorescence detection reagent
  • Adenosine triphosphate (ATP) fluorescence detection reagent

Examples

Experimental program
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Embodiment 1

[0020] The invention provides a kind of ATP fluorescence detection reagent, in the enzyme reaction basic solution system and the buffer system, a protective agent for improving the luciferase thermal stability in the enzyme reaction basic solution system is added, and the protective agent includes a concentration of 0.1-10M trehalose and 0.1-10M betaine. When the protective agent includes both betaine and trehalose, the ratio of betaine and trehalose is the same, and the concentration of betaine is 0.18-0.36M, and the concentration of trehalose and betaine is 0.62-1.24M.

[0021] The ATP fluorescence detection reagent consists of an enzyme reaction basic solution system, a buffer system and a protective agent.

[0022] Wherein, the buffer system is a trimethylglycine buffer solution, and the concentration of the trimethylglycine buffer solution is 0.1M.

[0023] The enzyme reaction basic solution system includes luciferase, luciferin and metal ions, the luciferase is preferab...

Embodiment 2

[0041] Compared with Example 1, the difference between the configuration method of the ATP fluorescence detection reagent prepared in Example 1 and the verification method of fluorescence value in this example is:

[0042] The storage temperature of the ATP fluorescence detection reagent in step S5 is 35°C;

[0043] In step S6, on day 0, day 1, day 2, day 4, day 7, day 8, day 10 and day 14, the ATP fluorescence detection reagent subpackaged in step S5 is taken out, put into a fluorescence detector and swabbed. Add 100uL of 0.013g / L ATP to the sub-tube, mix well, and test the fluorescence value.

[0044] Please refer to Figure 4 , Figure 4 It is the fluorescence value curve graph of adding different amounts of trehalose and betaine to the ATP fluorescence detection reagent under the condition of 35°C, wherein:

[0045] Curve 1 is the fluorescence numerical curve of 750 μL blank group without adding trehalose and betaine;

[0046] Curve 2 is the fluorescence numerical curv...

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Abstract

The invention provides an adenosine triphosphate (ATP) fluorescence detection reagent. According to the ATP fluorescence detection reagent, protective agents for improving luciferase thermal stabilityin an enzyme reaction basic solution system are added into the enzyme reaction basic solution system and a buffer system, and the protective agents comprise trehalose and glycine betaine with specific concentration. Compared with related technologies, the thermal stability of the ATP fluorescence detection reagent is better and retention time is obviously prolonged.

Description

【Technical field】 [0001] The invention relates to the technical field of ATP fluorescence detection, in particular to an ATP fluorescence detection reagent stable at room temperature. 【Background technique】 [0002] In the presence of ATP (adenosine triphosphate, adenosine triphosphate), luciferase oxidizes luciferin and emits fluorescence, and the emitted photons can be detected by light-sensitive elements, such as fluorescent detectors or improved optical microscopes . [0003] Luciferase has poor thermal stability and loses activity quickly at room temperature. Increasing the thermostability of luciferase can be carried out by changing its gene structure (gene mutation) on the one hand. But this method is cumbersome, and many mutations that can change its thermal stability have been patented. Another method is to add specific protective agents to improve the stability of luciferase. [0004] Although various commercially available ATP fluorescence detection reagents h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/26G01N21/64
CPCC12Q1/26G01N21/6486
Inventor 李书红陈杨青夏放蒋晓东
Owner 浙江东鸿生物科技有限公司