Method for determining content of soybean antigen protein

A soybean antigen protein and assay method technology, which is applied in the field of biological detection, can solve the problems of low reliability, complicated operation, low detection accuracy, etc. Suitable for promotion and application

Inactive Publication Date: 2019-06-11
TIANJIN RUIFU FEED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The embodiment of the present invention provides a method for determining the content of soybean antigenic protein, which aims to solve the technical problem

Method used

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  • Method for determining content of soybean antigen protein
  • Method for determining content of soybean antigen protein

Examples

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Embodiment 1

[0031] A method for assaying soybean antigen protein content, comprising the following steps:

[0032] Prepare a phosphate solution (0.05mol / L) containing sodium sulfite (0.02mol / L) as a buffer;

[0033] Crush 5g of soybean meal sample to 50 mesh, mix it with the buffer solution at a material-to-liquid ratio of 1:25, and perform water bath treatment (water bath treatment temperature is 35°C, time is 1h), take it out and shake well, and obtain the first mixed liquid;

[0034] Adding dithiothreitol (0.01mol / L) and sodium lauryl sulfate (the volume ratio of the first mixed solution to dithiothreitol is 100: 1) Slowly raise the temperature to 40°C at 1°C / min, and treat it in a constant temperature water bath for 2-3 hours, and shake it once every 5 minutes, and let it stand to cool to obtain the second mixed solution;

[0035] Centrifuge the second mixed solution, and take the filtrate;

[0036] Adjust the pH of the filtrate to 6.1, and centrifuge at 2°C, and the resulting prec...

Embodiment 2

[0040] A method for assaying soybean antigen protein content, comprising the following steps:

[0041] Prepare a phosphate solution (0.06mol / L) containing sodium sulfite (0.03mol / L) as a buffer;

[0042] Crush 5g of soybean meal sample to 60 mesh, mix it with the buffer solution at a material-to-liquid ratio of 1:30, and perform water bath treatment (water bath treatment temperature is 40°C, time is 1.5h), take it out and shake well, and obtain the first mixed solution;

[0043] Adding dithiothreitol (0.02mol / L) and sodium lauryl sulfate (the volume ratio of the first mixed solution to dithiothreitol is 100: 1) Slowly raise the temperature to 45°C, and treat it in a constant temperature water bath for 2.5 hours, and shake it every 8 minutes, and let it cool down to obtain the second mixed solution;

[0044] Centrifuge the second mixed solution, and take the filtrate;

[0045] Adjust the pH of the filtrate to 6.2, and centrifuge at 3°C, the resulting precipitate is 11S globuli...

Embodiment 3

[0049] A method for assaying soybean antigen protein content, comprising the following steps:

[0050] Prepare a phosphate solution (0.07mol / L) containing sodium sulfite (0.03mol / L) as a buffer;

[0051] Crush 5g of soybean meal sample to 70 mesh, mix it with the buffer solution at a material-to-liquid ratio of 1:35, and perform water bath treatment (water bath treatment temperature is 45°C, time is 2h), take it out and shake well, and obtain the first mixed mixture. liquid;

[0052] Adding dithiothreitol (0.03mol / L) and sodium lauryl sulfate (the volume ratio of the first mixed solution to dithiothreitol is 100: 1) Slowly raise the temperature to 50°C, and treat it in a constant temperature water bath for 3 hours, shake it up every 10 minutes, let it stand and cool down, and get the second mixed solution;

[0053] Centrifuge the second mixed solution, and take the filtrate;

[0054] The pH of the filtrate was adjusted to 6.2, and centrifuged at 5°C, the resulting precipita...

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Abstract

The invention discloses is applicable to the technical field of biological detection, and provides a method for determining content of soybean antigen protein. The method comprises the following stepsof preparing a phosphate solution containing sodium sulfite as a buffer solution; mixing a soybean meal sample with the buffer solution according to a certain ratio in order to obtain a first mixed solution; adding a certain proportion of dithiothreitol and sodium dodecyl sulfate to the first mixed solution, slowly heating to 40 to 50 DEG C, and carrying out constant-temperature water bath treatment for 2 to 3 hours in order to obtain a second mixed solution; carrying out centrifugal treatment on the second mixed solution, and taking a filtrate; adjusting pH of the filtrate to be 6.1 to 6.2,wherein the obtained precipitate is 11S globulin; adjusting the pH of the filtrate after the precipitation to be 3.8 to 3.9, wherein the obtained precipitate is 7S globulin; and determining the content of 11S globulin and 7S globulin. The method is convenient and rapid, the antigen protein can be separated within a short time without expensive instrument equipment, the sensitivity is high, the influence from external factors is small, and the method is suitable for popularization and application.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a method for measuring the content of soybean antigen protein. Background technique [0002] The nutrients contained in soybean are relatively comprehensive, rich in high-quality protein, and the balance of amino acid composition is good. It is considered as a high-quality plant-based protein resource and is widely used in animal feed. However, some anti-nutritional factors in it will affect animals. Especially the negative effects of sensitization in young animals, causing a series of adverse reactions such as allergic diarrhea and intestinal mucosal cell proliferation in young animals, and even death. Most of these antinutritional factors are antigenic proteins, so the research progress of isolating antigenic proteins from soybean is more and more abundant, so there are more requirements for the judgment of the isolation of antigenic proteins. [0003] The qualita...

Claims

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Application Information

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IPC IPC(8): G01N31/02
Inventor 赵蕊
Owner TIANJIN RUIFU FEED CO LTD
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