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
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Embodiment 1
[0059] Embodiment 1: Novel absorption system detects the H released by the NaHS donor 2 S content
[0060] It has been shown in the literature 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 pool, and the experimental group added the same volume of different concentrations of NaHS donors to the reaction pool. 2 S fluorescent probe working solution), shake lightly for 1 hour at 37°C, use the reaction bottle-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 as figure 2 It was shown that the fluorescence intensity of the high concentration group (600 μmol / L) was greater than that of the low concentration group (200 μmol / L), and the fluorescence intensity of the experimental group was significantly greater than that of the control group.
Embodiment 2
[0061] Example 2: Different absorption times, H 2 The S fluorescent probe detects the H released from the NaHS donor 2 S content
[0062] The control group added a certain volume of PBS to the reaction pool, and the experimental group added the same volume of NaHS donors with different concentrations to the reaction pool. 2 S fluorescent probe working solution), at 37°C, shake gently for different times, and use the reaction bottle-fluorescent probe method to detect the fluorescence intensity, which can reflect the H in the absorption solution 2 S content. The result is as image 3 It shows 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 intensity of the 1h group is slightly greater than that of the 1 / 2h group. ** P## P<0.0...
Embodiment 3
[0063] Example 3: H 2 The S fluorescent probe directly detects the H released from the NaHS donor 2 S content
[0064] Prepare different concentrations of NaHS donors, 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 as Figure 4 display, with the H 2 As the concentration of S increased, the fluorescence intensity increased correspondingly in a dose-dependent manner. ** P## P<0.01 compared with 0 μmol / L NaHS group.
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