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A method for identifying silk species from ancient silk fabric fragments based on immunotrace method

A technology for silk fabrics and fragments is applied in the field of cultural relic detection, which can solve the problems of large influence of impurities and low technical sensitivity, and achieve the effects of strong fluorescence stability, full utilization of resources and pollution reduction.

Active Publication Date: 2019-03-12
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

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
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1) Preparation of fluorescent carbon dots (CDs): Weigh 2.00g sweet potato tuber fragments and 6.00g KH 2 PO 4 , add 10mL of distilled water, pour the mixed solution into a hydrothermal reaction kettle and seal it, place it in an oven at 180°C and heat it for 10 h, after cooling to room temperature, take out the reaction solution, sonicate it for 10 min, and filter it with a 0.22 μm microporous membrane to obtain a shallow Yellow fluorescent carbon dot liquid; dialyze the obtained solution with a dialysis bag (molecular weight cut-off 1000) for 24 h to remove other small molecular impurities; finally freeze-dry the purified carbon dot solution to obtain fluorescent carbon dot powder.

[0044] 2) Preparation of fluorescent carbon dot-labeled silk fibroin secondary antibody: add fluorescent carbon dots, carbodiimide and secondary antibody to 10 mmol / L PBS buffer, stir at room temperature for 2-3 h, then use The PBS buffer was used to wash and centrifuge the solution sever...

Embodiment 2

[0057] 1) Preparation of fluorescent carbon dots (CDs): Weigh 2.00g sweet potato tuber fragments and 6.00g KH 2 PO 4 , add 10 mL of distilled water, pour the mixed solution into a hydrothermal reaction kettle and seal it, place it in an oven at 200°C and heat it for 13 h, after cooling to room temperature, take out the reaction solution, sonicate it for 10 min, and filter it with a 0.22 μm microporous membrane to obtain a shallow Yellow fluorescent carbon dot liquid; dialyze the obtained solution with a dialysis bag (molecular weight cut-off 1000) for 24 h to remove other small molecular impurities; finally freeze-dry the purified carbon dot solution to obtain fluorescent carbon dot powder.

[0058] 2) Preparation of fluorescent carbon dot-labeled silk fibroin secondary antibody: add fluorescent carbon dots, carbodiimide and secondary antibody to 10 mmol / L PBS buffer, stir at room temperature for 2.5 h, then wash with PBS buffer The solution is washed and centrifuged several ...

Embodiment 3

[0071] 1) Preparation of fluorescent carbon dots (CDs): Weigh 2.00g sweet potato tuber fragments and 6.00g KH 2 PO 4 , add 10 mL of distilled water, pour the mixed solution into a hydrothermal reaction kettle and seal it, place it in an oven at 220°C and heat it for 15 h, after cooling to room temperature, take out the reaction solution, ultrasonicate for 10 min, and filter it with a 0.22 μm microporous membrane to obtain a shallow Yellow fluorescent carbon dot liquid; dialyze the obtained solution with a dialysis bag (molecular weight cut-off 1000) for 24 h to remove other small molecular impurities; finally freeze-dry the purified carbon dot solution to obtain fluorescent carbon dot powder.

[0072] 2) Preparation of fluorescent carbon dot-labeled silk fibroin secondary antibody: add fluorescent carbon dots, carbodiimide and secondary antibody to 10 mmol / L PBS buffer, stir at room temperature for 3 h, then wash with PBS buffer The solution is washed and centrifuged several ...

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
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Abstract

The invention relates to the field of detection of cultural relics and discloses a method for identifying the varieties of residual silk of antique silk fabric based on an immune trace method. The method comprises the following steps: preparing a fluorescent carbon point by taking a sweet potato as a carbon source, marking secondary antibodies of mulberry silk and tussah silk by using the fluorescent carbon point, hydrolyzing promiscuous antique silk fabric cultural relic samples by using ionic liquid and PM13-alkaline protease step by step, performing dialysis and purification after protein extracting liquid is obtained, performing SDS-PAGE gel electrophoresis, transferring the obtained protein band to a PVDF membrane, incubating silk primary antibodies and the secondary antibodies marked by the fluorescent carbon point, observing the immunofluorescence band in a gel imaging system, and identifying the varieties of the antique silk fabric. According to the method, the use amount of a chemical reagent is small, and the reaction is mild, environment-friendly and harmless; and when the antique silk fabric is detected, the method has the characteristics of small use amount of samples, intuitive performance, rapidness and high sensitivity.

Description

technical field [0001] The invention relates to the field of detection of cultural relics, in particular to a method for identifying silk types of ancient silk fabric fragments based on an immunological trace method. Background technique [0002] For thousands of years, silk has always been a symbol of ancient China, and the long and splendid Chinese culture spread to the world through the Silk Road. Silk is a natural protein fiber, which is a natural polymer material connected by amino acids through peptide bonds. Commonly used silks include mulberry silk, tussah silk and castor silk, etc. Because of their different amino acid content and intermolecular microstructure, their textiles have different mechanical properties and wearability. [0003] Among the ancient silk remains, it can be seen from the archaeological excavations over the years that the manifestations of silk remains are mainly divided into four types: silk fabrics, gray silk fabrics, mineralized silk fabrics...

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
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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/533
CPCG01N33/533G01N33/6803
Inventor 胡智文陈茹茹李津梁军龙郑浩然
Owner ZHEJIANG SCI-TECH UNIV
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