Fluorescent assay method for urease activity based on gold nanoclusters
A gold nanocluster and urease activity technology, applied in analytical chemistry and nanometer fields, can solve problems such as environmental pollution and economic loss, and achieve the effects of high detection sensitivity and simple and fast preparation process.
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example 1
[0033] Add 0.6 mL of 0.5 mol / L sodium hydroxide solution and 0.4 mL of 0.02 g / L chloroauric acid solution to 4 mL of 0.08 mol / L N-acetyl-L-cysteine solution Mix well and place in a constant temperature water bath at 37°C for 2.5 hours. After the reaction, the reaction solution was purified with a dialysis bag with a molecular weight cutoff of 3500. The obtained gold nanocluster solution is colorless under visible light, and produces strong red fluorescence under ultraviolet light irradiation. Stored in the dark at 4°C, it can remain relatively stable for at least one month.
example 2
[0035] Add 0.05 mL of 22 U / L urease solution (pH=6.0) to 0.2 mL of 1 mol / L urea solution (pH=6.0), mix well and react in a constant temperature water bath at 25°C 40 min. Add 0.2 mL of the gold nanocluster solution prepared in Example 1 to the above solution, and react in a constant temperature water bath at 25°C for 3 min. Set a group of blank control group without urease. After the reaction, observe under ultraviolet light, the blank control group shows red fluorescence ( figure 1 A), and the red fluorescence of gold nanoclusters added with 22 U / L urease is quenched ( figure 1 In B). figure 2 The fluorescence emission spectra of the gold nanocluster solution of the blank control group and the urease group.
example 3
[0037] Add 0.05 mL of 22 U / L urease solution (pH=6.0) to 0.2 mL of different concentrations of urea solution (pH=6.0), mix well and react in a constant temperature water bath at 25°C for 40 min. Add 0.2 mL of the gold nanocluster solution prepared in Example 1 to the above solution, react in a constant temperature water bath at 25°C for 3 minutes, and measure the emitted light intensity F 650 . by image 3 It can be seen that when the urea concentration is 1mol / L, the fluorescence quenching value ΔF 650 to reach maximum.
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