Novel method for detecting activity of hydrogen sulfide synthetase by using hydrogen sulfide fluorescence probe and application of method
A technology for fluorescent probes and detection methods, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve problems such as cumbersome steps, and achieve the effects of good repeatability, good stability, and high quantum yield
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[0059] Example 1: A new absorption system detects H released by NaHS donor 2 S content
[0060] The literature shows that HS in NaHS solution - / H2S ratio is 3:1, so most of the NaHS solution is HS - . The control group added a certain volume of PBS to the reaction tank, and the experimental group added the same volume of different concentrations of NaHS donors to the reaction tank. Both groups added the same absorption system (containing the reaction adjuvant H 2 S fluorescent probe working solution), 37℃, gently shake for 1 hour, use the reaction flask-fluorescent probe method to detect the fluorescence intensity, the fluorescence intensity can reflect the H in the absorption solution 2 S content. The result is figure 2 It shows that the fluorescence intensity of the high concentration group (600μmol / L) is greater than that of the low concentration group (200μmol / L), and the fluorescence intensity of the experimental group is significantly greater than that of the control group...
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[0061] Example 2: Different absorption time, H 2 S fluorescent probe detects H released by NaHS donor 2 S content
[0062] The control group added a certain volume of PBS to the reaction tank, and the experimental group added the same volume of NaHS donors of different concentrations to the reaction tank. Both groups added the same absorption system (containing the reaction adjuvant alkanium bromide H 2 S fluorescent probe working solution), 37℃, gently shake for different time, use the reaction flask-fluorescent probe method to detect the fluorescence intensity, the fluorescence intensity can reflect the H in the absorption solution 2 S content. The result is image 3 It is shown that the fluorescence intensity of the 600μmol / L NaHS group is greater than that of the 200μmol / L NaHS group, and the fluorescence intensity of the experimental group is significantly greater than that of the control group. In the experimental group, with the same NaHS concentration, the fluorescence inte...
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[0063] Example 3: H 2 S fluorescent probe directly detects H released by NaHS donor 2 S content
[0064] Prepare different concentrations of NaHS donors separately and add the same H 2 S fluorescent probe, using a full-wavelength scanning multifunctional reader (Varioskan Flash3001) (maximum excitation wavelength λex(max)=476nm, maximum emission wavelength λem(max)=513nm). The result is Figure 4 Show that with H 2 As the concentration of S increases, the fluorescence intensity increases accordingly, in a dose-dependent manner. ** P## P<0.01 compared with 0μmol / L NaHS group.
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