A new method for detecting the activity of hydrogen sulfide synthase with a fluorescent probe of hydrogen sulfide and its application

A technology of fluorescent probes and detection methods, which is applied in the fields of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of cumbersome steps, and achieve the effects of good repeatability, thorough cracking degree and good stability
CN103630518BInactive Publication Date: 2017-11-07蔡典其 +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
蔡典其
Publication Date
2017-11-07
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the technical field of biological detection, relates to a new method for detecting the activity of hydrogen sulfide synthase with a fluorescent probe of hydrogen sulfide, and provides an application path of the method. It is suitable for detecting the activity of hydrogen sulfide synthase including CBS, CSE and CL. For tissue samples, first cut into small pieces and then grind. For cells, tissue cell lysate is used for lysing, and the lysate is used for protein quantification. As shown in the figure, the reaction system ( Tissue cell lysates, L-cysteine, 5-pyridoxal phosphate) were added to the reaction pool on the outer ring of the reaction flask, and the absorption system (H2S fluorescent probe working solution containing reaction adjuvant alkane ammonium bromide) was added to In the absorption pool in the inner ring, the H2S gas molecules produced by the reaction system are combined with the fluorescent probes of the absorption system, which can be used for fluorescence measurement and fluorescence imaging, quantitatively detect the H2S content, and more intuitively reflect the activity of H2S synthase. The invention can provide a good research method for studying H2S-related signaling pathways and the significance and diagnosis of H2S in some disease processes.
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Description

1. Technical field

[0001] The invention belongs to the technical field of biological detection, and in particular relates to a new method for detecting the activity of hydrogen sulfide synthase with a hydrogen sulfide fluorescent probe and an application thereof. This method of detecting enzyme activity with fluorescent probes can be used to study the mechanism of some important enzymatic processes in tissue cells, clarify some important signaling pathways in biological systems, and provide good assistance for the early diagnosis and prevention of some major diseases means. Two background technology

[0002] For a long time, hydrogen sulfide (H 2 S) is considered a toxic gas, however, with the 2 With the in-depth study of S, it is found that some tissue cells in the body can produce H 2 S, it is the third endogenous gas signal molecule discovered after nitric oxide (NO) and carbon monoxide (CO). More and more experiments show that H 2 S is not only an endogenous gas sig...

Claims

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