Method for quickly extracting, purifying and detecting protein in hair

A protein and hair sample technology, applied in the field of rapid extraction, purification and detection of hair sample protein, can solve the problems of unfavorable protein composition analysis, low yield, harmful to human body, etc., achieve reactivity, easy solubility, low toxicity, and easy operation Effect

Inactive Publication Date: 2014-10-15
NORTHWEST A & F UNIV
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Problems solved by technology

The mechanical method can not only break the disulfide bond, but also break the polypeptide chain. The yield is not high, which is not conducive to the analysis of protein components.
During the acid-base extraction process in the chemical method, alkaline waste water and waste acid steam will be generated, which will pollute the environment and be harmful to the human body

Method used

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  • Method for quickly extracting, purifying and detecting protein in hair

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Embodiment Construction

[0026] The technical solution of the present invention will be further described in detail below in conjunction with the drawings and specific embodiments.

[0027] A method for rapid extraction, purification and detection of hair-like protein, including the following steps:

[0028] (1) Samples are classified and cut into small sections respectively;

[0029] (2) 0.15% nonylphenol polyoxyethylene (9) ether, cleaned at 60°C for 20 minutes, and sucked dry;

[0030] (3) Rinse with dd water at 60°C for 5 minutes, then blot dry;

[0031] (4) Wash with dichloromethane and ethanol, wash at room temperature for 20 minutes, and blot dry;

[0032] (5) Rinse with ultrapure water at 60°C for 5 minutes, and then absorb dry;

[0033] (6) 0.1M methanol base KOH, room temperature for 30 minutes, suck dry;

[0034] (7) Rinse with ultrapure water at 60°C for 5 minutes, and then absorb dry;

[0035] (8) Grinding: soak the plush with water and grind it in liquid nitrogen;

[0036] (9) Extraction: Take 200mg of...

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Abstract

The invention discloses a method for quickly extracting, purifying and detecting protein in hair. The method is characterized by comprising the following steps: (1) classifying samples and respectively shearing the samples into small sections; (2) cleaning for 20 min with 0.15 percent nonylphenol polyoxyethylene ether (9) at the temperature of 60 DEG C and then sipping up; (3) cleaning for 5 min with dd water at the temperature of 60 DEG C and then sipping up; (4) cleaning for 20 min with methylene dichloride and ethanol at the room temperature and then sipping up; (5) cleaning for 5 min with the dd water at the temperature of 60 DEG C and then sipping up; (6) cleaning for 30 min with 0.1 M KOH at the room temperature and then sipping up; (7) cleaning for 5 min with the dd water at the temperature of 60 DEG C and then sipping up; (8) grinding with liquid nitrogen after water soaking; (9) extracting: overnight slaking after adding 7M urea of 200 [mu] L, 2M , 50mM Tris and 50mM TCEP (pH4) at the room temperature; (10) purifying.

Description

Technical field [0001] The invention belongs to the technical field of chemistry and relates to a method for rapid extraction, purification and detection of hair-like protein. Background technique [0002] Hair protein is mainly composed of keratin, and keratin molecules mainly rely on disulfide bonds to form a stable spatial network structure. Dissolving the protein in the hair is mainly to break the disulfide bonds (-S-S). In addition, the high-density hydrogen bonds and salt bonds between protein molecules cannot be ignored. At present, the current domestic protein extraction technology takes a relatively long time and the procedures are cumbersome, which is not conducive to the high-throughput and large-scale application of the laboratory. The main methods for preparing keratin are as follows: 1) Mechanical methods, including high-pressure hydrolysis, expansion, Extrusion method and extrusion method. The main method is heating (100-200°C) and pressure (0.294-0.98MPa) to bre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/34
Inventor 王小龙陈玉林周光现牛怡源师丙波
Owner NORTHWEST A & F UNIV
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