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A New Method for Identifying Pearl Protein Components

A technology of pearl protein and a new method, which is applied in the field of biochemical analysis, can solve problems such as unsuitable identification of pearl protein components, and achieve the effects of increasing migration distance, improving solubility, and reducing resolution loss

Active Publication Date: 2020-06-16
JINHUA VOCATIONAL TECH COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is aimed at large molecular weight proteins with protein molecular weights above 100kDa, and the molecular weight distribution of total pearl proteins is wide, so this method is not suitable for identifying pearl protein components.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A new method for identifying pearl protein components, including electrophoresis detection, liquid chromatography separation, mass spectrometry detection and protein database analysis, the specific steps are:

[0021] 1) Electrophoresis separation: use 10% sodium dodecylsulfonate-polyacrylamide separation gel to carry out SDS-PAGE electrophoresis on the pretreated pearl protein, 80V constant voltage for 15min, 120V constant voltage to the end point, and gel after electrophoresis Stain with Coomassie Brilliant Blue R-250 for 4 hours, then decolorize until the background is clear, then scan the spectrum and analyze the bands. The above separation gel contains 0.01‰ graphene, and the weight ratio of reduced graphene and graphene oxide in graphene is 1:4.7, the carboxyl ratio of graphene oxide is 2wt%;

[0022] 2) Liquid chromatography separation: cut off the colored protein gel, wash it, dry it, pretreat it with dithiothreitol as a reducing agent at 60°C for 50 minutes, an...

Embodiment 2

[0027] A new method for identifying pearl protein components, the concrete steps are:

[0028] 1) Electrophoresis separation: use 15% sodium dodecylsulfonate-polyacrylamide separation gel to carry out SDS-PAGE electrophoresis on the pretreated pearl protein, 80V constant voltage for 15min, 120V constant voltage to the end point, and gel after electrophoresis Stain with Coomassie Brilliant Blue R-250 for 2 hours, then decolorize until the background is clear, then scan the spectrum and analyze the bands. The above separation gel contains 0.02‰ graphene, and the weight ratio of reduced graphene and graphene oxide in graphene is 1:4.3, the carboxyl ratio of graphene oxide is 4wt%;

[0029] 2) Liquid chromatography separation: Cut off the colored protein gel, wash it, dry it, pretreat it with reducing agent dithiothreitol at 50°C for 70 minutes, and then treat it with iodoacetamide for 40 minutes in the dark room , take it out, put it into trypsin solution, treat at 5°C for 20min...

Embodiment 3

[0034] A new method for identifying pearl protein components, the concrete steps are:

[0035] 1) Electrophoresis separation: use 12% sodium dodecylsulfonate-polyacrylamide separation gel to carry out SDS-PAGE electrophoresis on the pretreated pearl protein, 80V constant voltage for 15min, 120V constant voltage to the end point, and gel after electrophoresis Stain with Coomassie Brilliant Blue R-250 for 3 hours, then decolorize until the background is clear, then scan the spectrum and analyze the bands. The above separation gel contains 0.05‰ graphene, and the weight ratio of reduced graphene and graphene oxide in graphene is 1:4.5, the carboxyl ratio of graphene oxide is 3wt%;

[0036] 2) Liquid chromatography separation: cut off the colored protein gel, wash it, dry it, pretreat it with reducing agent dithiothreitol at 55°C for 60 minutes, and then treat it with iodoacetamide for 45 minutes in the dark room , put it into trypsin solution after taking it out, treat it at 3°C...

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Abstract

The invention discloses a new method for identifying the pearl protein component. The new method comprises the steps that pearl protein is subjected to SDS-PAGE, after electrophoresis is conducted, gel is dyed with coomassie brilliant blue, decoloration is conducted, spectrum scanning is conducted, and a strip is analyzed; protein gel obtained after color development is cut off, washed to be clean, dried, pretreated with dithiothreitol, treated with iodoacetamide and then treated with trypsin, and finally peptide is separated and extracted with liquid chromatogram for use; the peptide is subjected to mass spectrometric detection, and a mass spectrum is obtained; the mass spectrum is subjected to database analysis. The method has the advantages that operation is easy, rapid and efficient, acarbohydrate chain in the pearl protein can be removed, the analysis difficulty can be lowered, the component of soluble protein in pearl can be analyzed and identified, and the theoretical basis isprovided for further deep research and comprehensive development of the pearl protein.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis, in particular to a new method for identifying pearl protein components. Background technique [0002] Hyriopsiscumingii Lea is a species endemic to my country, belonging to the phylum Mollusca, class Lamellibranchia, order Eulamellibranchia, family Unionidae, genus Hypriosis, and is a freshwater benthic mollusc. , Widely distributed in Poyang Lake, Dongting Lake, Taihu Lake and other large and medium-sized lakes and their watersheds. The pearls it produces are fine and smooth in texture, bright in color and crystal clear, and are the main varieties of freshwater pearls produced in my country. The color of pearls has basic hues such as white, yellow, purple, red, and black. Each hue contains many chromaticities, and the uniformity of color and chromaticity is one of the important indicators to measure the value of pearls. In order to make consistent colors, chromaticity Uniform high-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447G01N30/02G01N33/68
CPCG01N27/447G01N30/02G01N33/6848
Inventor 叶容晖
Owner JINHUA VOCATIONAL TECH COLLEGE
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