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Method for detecting ancient argillic silk fabric on basis of immunoblotting

A technology of silk fabrics and mudification, which is applied in the field of detection of ancient muddy silk fabrics based on the immune trace method, can solve the problems of great influence of impurities, unsuitability for detection of trace cultural relic samples, low technical sensitivity, etc., achieve strong fluorescence stability, reduce silk The effect of protein loss and full utilization of resources

Active Publication Date: 2017-11-24
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the present stage of research, most of the research methods on ancient silk fabrics at home and abroad still use infrared spectroscopy, Raman spectroscopy, X-ray diffraction and other techniques. These techniques have low sensitivity and are greatly affected by impurities, so they are not suitable for the detection of trace cultural relic samples. , how to find a more scientific method to identify ancient silk fabrics has always been a difficult problem in the field of cultural relics protection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1) Preparation of fluorescent carbon dots (CDs): Measure 10 mL of deionized water and absolute ethanol (volume ratio 1:1), mix and dissolve in a beaker, add 0.40 g fructose, and stir until completely dissolved. Pour the mixed solution into a hydrothermal reactor and seal it, place it in an oven at 160℃ and heat it for 2 hours. When the solution turns to light yellow and the reaction is complete, cool to room temperature naturally to obtain a fluorescent carbon dot solution; use a dialysis bag for the obtained solution (Molecular weight cut-off: 1000) Dialysis for 24 hours to remove other small molecule impurities; finally, the purified carbon dot solution is freeze-dried to obtain fluorescent carbon dot powder.

[0046] 2) Preparation of fluorescent carbon-dot-labeled silk fibroin secondary antibody: add fluorescent carbon-dot, carbodiimide and secondary antibody into 10mmol / L PBS buffer at a mass ratio of 1:20:20 at room temperature After stirring for 2 h, the solution wa...

Embodiment 2

[0059] 1) Preparation of fluorescent carbon dots (CDs): Measure 10 mL of deionized water and absolute ethanol (volume ratio 1:1), mix and dissolve in a beaker, add 0.40 g of fructose, and stir until completely dissolved. Pour the mixed solution into a hydrothermal reactor and seal it, place it in a 160℃ oven and heat it for 2.5 hours. When the solution turns to light yellow and the reaction is complete, cool to room temperature naturally to obtain a fluorescent carbon dot solution; use a dialysis bag for the obtained solution (Molecular weight cut-off: 1000) Dialysis for 24 h to remove other small molecule impurities; finally, the purified carbon dot solution is freeze-dried to obtain fluorescent carbon dot powder.

[0060] 2) Preparation of fluorescent carbon-dot-labeled silk fibroin secondary antibody: add fluorescent carbon-dot, carbodiimide and secondary antibody into 10mmol / L PBS buffer at a mass ratio of 1:20:20 at room temperature After stirring for 2.5 h, the solution was...

Embodiment 3

[0073] 1) Preparation of fluorescent carbon dots (CDs): Measure 10 mL of deionized water and absolute ethanol (volume ratio 1:1), mix and dissolve in a beaker, add 0.40 g of fructose, and stir until completely dissolved. Pour the mixed solution into a hydrothermal reaction kettle and seal it, place it in a 160℃ oven and heat it for 3 hours. When the solution turns light yellow and the reaction is complete, cool to room temperature naturally to obtain a fluorescent carbon dot solution; use a dialysis bag for the obtained solution (Molecular weight cut-off: 1000) Dialysis for 24 h to remove other small molecule impurities; finally, the purified carbon dot solution is freeze-dried to obtain fluorescent carbon dot powder.

[0074] 2) Preparation of fluorescent carbon-dot-labeled silk fibroin secondary antibody: add fluorescent carbon-dot, carbodiimide and secondary antibody into 10mmol / L PBS buffer at a mass ratio of 1:20:20 at room temperature After stirring for 2~3 h, the solution ...

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Abstract

The invention relates to the field of cultural relic detection and discloses a method for detecting ancient argillic silk fabric on the basis of immunoblotting. The method comprises steps as follows: firstly, a silk secondary antibody marked with fluorescent carbon points is prepared, a modern silk sample and an ancient argillic silk fabric sample are hydrolyzed with an ionic liquid and PM13-alkaline protease step by step, a protein extraction solution is obtained and subjected to dialysis, purification and SDS-PAGE gel electrophoresis, an obtained protein band is transferred to a PVDF (polyvinylidene fluoride) film and subjected to incubation with a silk first antibody and the silk secondary antibody marked with the fluorescent carbon points, an immunofluorescence band can be observed in a gel imaging system, and the type of ancient silk is identified. The consumption of chemical reagents is low, the reaction is mild, and the method is environment-friendly and harmless; during detection for the ancient silk fabric, the method has the characteristics of being low in sample consumption, direct, rapid and high in sensitivity.

Description

Technical field [0001] The invention relates to the field of cultural relic detection, in particular to a method for detecting ancient muddy silk fabrics based on an immune trace method. Background technique [0002] my country is the origin of silk widely recognized in the world, and the invention of silk has a pivotal position in the history and culture of the Chinese nation. For thousands of years, silk has been a symbol of ancient China, and the long and splendid Chinese culture has spread to the world through the Silk Road. Chinese silk mainly uses silk, which is a natural protein fiber, which is a natural polymer material connected by amino acids through peptide bonds. Silk fabrics are buried under the ground, and protein fibers are easily aging and degraded under the influence of temperature, humidity, soil acidity and alkalinity, bacteria, molds, etc. [0003] In the ancient silk remains, from archaeological excavations over the years, it can be seen that the manifestati...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/533
CPCG01N21/6486G01N33/533
Inventor 胡智文陈茹茹李津梁军龙尚亚廷
Owner ZHEJIANG SCI-TECH UNIV
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