Method for detecting collagen in ancient cultural relic material

A technology of collagen and ancient cultural relics, which is applied in the detection field of collagen in ancient cultural relic materials, can solve the problems of inability to carry out efficient identification, and achieve the effects of low cost, simple detection and fast response

Active Publication Date: 2014-02-12
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are uncertain, especially when a collagen is derived from different materials or some protein molecular structures are similar, these methods cannot be efficiently identified at all.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] a) Solution preparation: preparation of PBS buffer solution: KCl 0.2g, KH 2 PO 4 0.2g, NaCl 8.0g, NaH 2 PO 4 ·7H 2 O 2.16g, dissolved in deionized water and made to a volume of 1000mL, adjusted to pH 7.4, sterilized at 121°C. Preparation of eluent: 5mL1mol / L trishydroxymethylaminomethane-HCl buffer solution, 1mL0.5mol / L ethylenediaminetetraacetic acid solution, 180g urea, 25mL20% sodium lauryl sulfate (50mL deionized water containing 10 g of solute), dissolved in deionized water and adjusted to 500 mL, adjusted to pH 7.4 with NaOH, and stored at room temperature after sterile suction filtration.

[0025] b) 100 μL of PBS buffer solution was set as a blank control, and 0.1 mg of fish gelatin powder was set as a positive control. Dissolve 100 μL of PBS buffer solution of blank sample, 0.1 mg of fish gelatin powder of positive control sample and 0.1 mg of cultural relic material of experimental sample in 100 μL of eluent respectively, and place it at room temperature...

Embodiment 2

[0041] A method for detecting collagen in ancient cultural relic materials, it comprises the steps:

[0042] a) Solution preparation: preparation of PBS buffer solution: KCl 0.2g, KH 2 PO 4 0.2g, NaCl 8.0g, NaH 2 PO 4 ·7H 2 O 2.16g, dissolved in deionized water and made to a volume of 1000mL, adjusted to pH 7.4, sterilized at 121°C. Preparation of eluent: 5mL1mol / L trishydroxymethylaminomethane-HCl buffer solution, 1mL0.5mol / L ethylenediaminetetraacetic acid solution, 180g urea, 25mL20% sodium lauryl sulfate (50mL deionized water containing 10 g of solute), dissolved in deionized water and adjusted to 500 mL, adjusted to pH 7.4 with NaOH, and stored at room temperature after sterile suction filtration.

[0043] b) 100 μL of PBS buffer solution was set as a blank control, and 0.1 mg of fish gelatin powder was set as a positive control. Dissolve 100 μL of blank sample PBS buffer solution, 0.1 mg of positive control sample fish gelatin powder and 0.3 mg of experimental sam...

Embodiment 3

[0059] A method for detecting collagen in ancient cultural relic materials, it comprises the steps:

[0060] a) Solution preparation: preparation of PBS buffer solution: KCl 0.2g, KH 2 PO 4 0.2g, NaCl 8.0g, NaH 2 PO 4 ·7H 2 O 2.16g, dissolved in deionized water and made to a volume of 1000mL, adjusted to pH 7.4, sterilized at 121°C. Preparation of eluent: 5mL1mol / L trishydroxymethylaminomethane-HCl buffer solution, 1mL0.5mol / L ethylenediaminetetraacetic acid solution, 180g urea, 25mL20% sodium lauryl sulfate (50mL deionized water containing 10 g of solute), dissolved in deionized water and adjusted to 500 mL, adjusted to pH 7.4 with NaOH, and stored at room temperature after sterile suction filtration.

[0061] b) 100 μL of PBS buffer solution was set as a blank control, and 0.1 mg of fish gelatin powder was set as a positive control. Dissolve 100 μL of PBS solution of blank sample, 0.1 mg of fish gelatin powder of positive control sample, and 0.5 mg of cultural relic m...

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PUM

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Abstract

The invention discloses a method for detecting collagen in an ancient cultural relic material. The method is characterized by comprising the following steps: detecting protein components in the ancient cultural relic material through a competitive enzyme-linked immunosorbent assay method, namely, fixing an AbCam rabbit anti-I type collagen polyclonal antibody on a solid phase carrier, and adding a sample eluent to form an antigen-antibody complex; washing through a phosphate buffer solution (PBS), and adding alkaline phosphatase-labeled goat anti-rabbit IgG(H+L) which is then bonded to the solid phase carrier through reaction, wherein the amount of enzyme on a solid phase is in a certain proportion to the quantity of detected substances in a sample; after a substrate of enzyme reaction is added, catalyzing the substrate into colored products through enzyme, wherein the quantity of the products is positively correlated to the quantity of the detected substances in the sample, so that qualitative analysis can be performed according to the color depth. The method disclosed by the invention is low in cost, simple in detection and high in response speed.

Description

technical field [0001] The invention relates to a method for detecting collagen in ancient cultural relic materials, which is mainly used for detecting collagen in ancient cultural relic materials. Background technique [0002] Cultural relics are the product of human social activities in a certain historical period and have important scientific value. Ancient cultural relic materials play a fundamental role in the study of cultural relics. Collagen, as a natural polymer material, is widely used in ancient cultural relic materials. In the past, there were many methods for identifying collagen in ancient cultural relics, mainly including microscope observation and identification, electron microscopy, gas chromatography, liquid chromatography, infrared spectrophotometry, and thermogravimetric analysis and identification. However, these methods are uncertain, especially when a collagen is derived from different materials or some protein molecular structures are similar, these...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/543G01N33/6803G01N2333/78
Inventor 刘苗苗郑益炜胡智文
Owner ZHEJIANG SCI-TECH UNIV
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