Method for visually detecting alkaline phosphatase

A phosphatase and alkaline technology, applied in the field of analysis and detection, can solve the problems that the detection method is susceptible to interference, etc., and achieve the effect of high sensitivity, simple detection process and simple steps

Inactive Publication Date: 2014-03-19
WUHAN UNIV
View PDF4 Cites 26 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because chloroauric acid has relatively strong oxidizing properties, it can be reduced even in weakly reducing solutions (such a

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for visually detecting alkaline phosphatase
  • Method for visually detecting alkaline phosphatase
  • Method for visually detecting alkaline phosphatase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The detection of embodiment 1 alkaline phosphatase

[0018] Taking alkaline phosphatase as an example, the method for visually detecting enzymes based on enzymatic metallation will be described in detail below.

[0019] 1. Method

[0020] 1. Preparation of gold nanoparticles

[0021] Weigh 7.5mg of glutathione, dissolve it in 0.1mL ultrapure water, add it into 1mL of 1% chloroauric acid solution under stirring, adjust the pH value to 2.5-3.0 with 10mol / L NaOH after mixing, and produce light yellow precipitation. The precipitate was collected by centrifugation (7000rpm, 3 minutes) to obtain the Au(I) complex.

[0022] The above Au(I) complex was dissolved with 1 mL of 5 mmol / L sodium hydroxide solution to obtain a gold precursor solution. Dilute the gold precursor solution with 9 mL of ultrapure water, and adjust the pH to around 5.5. Then, 4 mg of reduced coenzyme II (NADPH) and 2 units of glutathione reductase (GR) were added to the reaction system, and the reacti...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a method for visually detecting alkaline phosphatase. The method comprises the following steps: catalyzing a substrate p-aminophenol phosphate (p-APP) through the alkaline phosphatase to generate a substance p-aminophenol (p-AP) with strong reducing property; reducing Ag<+> into AgO through the p-AP in the presence of nanogold, and depositing the AgO on the surface of the nanogold to form Au/Ag nano-particles, so that the enzymatic activity can be detected according to the color change of a colloidal gold solution and the change of an ultraviolet-visible absorption spectrum. By the combination of enzymatic metallization and nanogold induced silver deposition, the ultrahigh-sensitivity method for visually detecting the alkaline phosphatase is established and has the characteristics of simplicity in operation, high specificity, high sensitivity, no need of a complicated instrument and the like.

Description

technical field [0001] The invention relates to a method for visually detecting enzyme activity, belonging to the field of analysis and detection. Background technique [0002] Visual detection is widely used in clinical diagnosis as an effective method for rapid detection due to its low requirements for instruments and simple operation. Colorimetric assays based on metal nanoparticles have attracted widespread attention due to their simplicity, convenience, and unique optical properties of metal nanoparticles. However, the colorimetric method based on the aggregation of metal nanoparticles still has some defects in practical applications, for example: metal nanoparticles are easily affected by some external factors, such as high ionic strength, complex substances in actual samples, etc. Non-specific aggregation of particles; in addition, the sensitivity of these detection methods needs to be further improved. [0003] Numerous types of enzymes have been reported, such as ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N21/78
Inventor 张志凌周川华赵静雅庞代文
Owner WUHAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products