An analysis method for detecting lipase activity based on gold nanocluster fluorescence enhancement
A technology of gold nano-clusters and analysis method, which is applied in the field of analysis of lipase activity based on fluorescence-enhanced detection of gold nano-clusters, and achieves the effects of simple and fast operation, low cost and simple method
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Embodiment 1
[0019] Example 1 Mechanism Verification
[0020] Add 800 μL of gold nanoclusters and 100 μL of PB buffer solution to 2 mL centrifuge tubes a, b, c, and d respectively, then add 100 μL of methyl thioglycolate to tube b, and add 100 μL of Methyl thioglycolate and 5 μg lipase, add 100 μL of methyl thioglycolate and 30 μg lipase to tube d, and make the other volume up to 1000 μL with ultrapure water, bathe in 35°C water for 8 minutes, and detect the fluorescence intensity (such as figure 1 shown).
[0021] figure 1 It is a mechanism verification diagram. It can be seen from curve a and curve b that the addition of the substrate methyl thioglycolate does not change the fluorescence intensity of gold nanoclusters. When 5 μg / mL lipase is added, the curve c, The fluorescence of gold nanoclusters is enhanced, and when 30 μg / mL lipase is added, the fluorescence intensity is further enhanced, indicating that the higher the concentration of lipase, the more the fluorescence intensity of...
Embodiment 2
[0022] The drafting of the standard curve of embodiment 2 lipase activity detection
[0023] Add 800 μL of gold nanoclusters, 100 μL of PB buffer solution, and 100 μL of methyl thioglycolate into a 2 mL centrifuge tube, bathe in water at 35°C for 8 min, and then add different concentrations of lipase, the concentration of lipase is as follows: 0 μmol / L , 5 μmol / L, 10 μmol / L, 15 μmol / L, 20 μmol / L, 25 μmol / L, 30 μmol / L, dilute to 1000 μL with ultrapure water, measure its fluorescence intensity after 35 ° C water bath for 8 min; the concentration of lipase is On the abscissa, the increased value of the fluorescence intensity is plotted on the ordinate, and the standard curve of lipase can be drawn (such as figure 2 shown).
[0024] figure 2 It shows that with the increase of the lipase concentration, the fluorescence intensity of the solution is gradually enhanced and the lipase concentration has a good linear relationship between 5-30μmol / L, and the detection limit is 0.37μg...
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