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Extraction method of goat milk fat globule membrane protein for proteome research

A milk fat globule membrane and proteomics technology, which is applied in the field of extraction of milk fat globule membrane protein in goat milk samples, can solve the problems of difficult extraction and low stability of goat milk fat globule membrane protein, and achieve stable properties. , The protein properties are stable, and the effect of improving the extraction rate

Inactive Publication Date: 2018-07-20
JILIN UNIV
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the existing proteomics as the research purpose, the protein is not easy to extract and the problem of low stability in the process of extracting goat milk fat globule membrane protein, and to provide a stable and capable Sample preparation and processing method for goat milk fat globule membrane proteomics research that effectively removes the influence of impurity components

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  • Extraction method of goat milk fat globule membrane protein for proteome research
  • Extraction method of goat milk fat globule membrane protein for proteome research

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Embodiment 1

[0023] Example 1: A method for extracting goat milk fat globule membrane protein for proteomics research

[0024] 1) Select fresh goat milk from healthy goats. In order to eliminate individual differences, select goat milk from 30 goats as whole milk samples. Take 50ml of goat milk from each goat. The fresh goat milk taken from the pasture must be at 4°C or -20°C as soon as possible. Transport it back to the laboratory for processing. If it cannot be processed in time, it should be stored at -80°C. Before use, it should be thawed in a refrigerator at 4°C. This method must use fresh or ultra-low temperature (-80°C) goat milk. 4 °C and -20°C are only suitable for short-term transportation, and cannot be used for storage of goat milk samples, otherwise protein degradation will occur and affect proteomics results.

[0025] 2) Add 4% (mass fraction) sucrose to the 30 goat milk samples in step 1), stir and mix with a magnetic stirrer to obtain a uniform goat milk sucrose solution. ...

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Abstract

The invention relates to an extraction method of a goat milk fat globule membrane protein for proteome research, and belongs to the technical field of milk fat globule membrane protein extraction. Themethod comprises the steps of adding cane sugar into a goat milk sample, performing high speed centrifugation to obtain crude separated milk fat, sequentially washing the crude extracted milk fat through a potassium phosphate buffer solution and ultrapure water, cracking through a lysis solution, and finally purifying through a precooled acetone solution, thus obtaining a milk fat globule membrane protein sample. According to the method provided by the invention, the influence of the low-content casein component can be effectively reduced, the extracted goat milk fat globule membrane proteinis higher in extraction rate and higher in stability, and the method has better popularization and application prospects and economic value in the goat milk fat globule membrane proteome research.

Description

technical field [0001] The invention relates to a method for extracting milk fat globule membrane protein for the purpose of proteomics analysis, in particular to an extraction method for milk fat globule membrane protein in a goat milk sample. Background technique [0002] Proteomics research is an in-depth and systematic study of the executive bodies of life activities-proteins at the level of proteomics, including protein expression levels, amino acid sequences, post-translational processing and protein interactions, and understanding of various aspects of cells at the protein level. Item functions, various physiological and biochemical processes, and pathological processes of diseases. [0003] The milk fat globule membrane is a thin film surrounding the milk fat globule, with a cross-sectional diameter of about 10-20nm, which acts as an emulsifier and prevents the polymerization and enzymatic degradation of the fat globule. There are 25% to 70% membrane proteins on the...

Claims

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Application Information

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IPC IPC(8): C07K14/47C07K1/14
CPCC07K14/47
Inventor 郭明若孙玉雪王翠娜
Owner JILIN UNIV
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