Method sutiable for two-dimensional electrophoresis extraction and separation of total protein of rat and mouse testicular tissue sample

A tissue sample and two-dimensional electrophoresis technology, which is applied in the field of protein extraction and proteomics separation, can solve the problems of time-consuming, labor-intensive, and high experimental costs, and achieve the effects of increasing solubility, good repeatability, and reducing time

Inactive Publication Date: 2013-09-04
SHANXI AGRI UNIV
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Problems solved by technology

The vast majority of samples need to explore the most suitable conditions through multiple

Method used

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  • Method sutiable for two-dimensional electrophoresis extraction and separation of total protein of rat and mouse testicular tissue sample
  • Method sutiable for two-dimensional electrophoresis extraction and separation of total protein of rat and mouse testicular tissue sample
  • Method sutiable for two-dimensional electrophoresis extraction and separation of total protein of rat and mouse testicular tissue sample

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Experimental program
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Embodiment Construction

[0026] Embodiments of the present invention are explained below.

[0027] The materials used in the present invention are: IPG prefabricated strips (Bio-Rad, USA), pH 4-7, 17cm, stored in -20°C refrigerator; carrier ampholyte (Bio-Rad, USA), stored in -4°C refrigerator; Focused loading buffer (Bio-Rad, USA), stored in refrigerator at 4°C; Urea, CHAPS, DTT (Dithiothreitol), Bromophenol Blue (Bromophenol Blue); Mineral Oil (Mineral Oil), SDS, Tris, Iodoacetamide; low melting point Agarose, Glycine, Acrylamide; Methylene Bisacrylamide (Bis), Ammonium Persulfate; TEMED, Glycerin, Thiuria, Bio-Safe TM Coomassie.

[0028] Instruments and software used in the present invention include: Y96000 type electronic balance (Germany Sartorius); T18 endotype tissue homogenizer (Germany IKA company); ultra-low temperature refrigerator (Thermo fisher); UV-2100 type spectrophotometer (especially Nika (Shanghai) Instrument Co., Ltd.); PROTEIN IEF System (BIO-Rad, USA); PROTEIN IEF xl Cell (BIO-...

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Abstract

The invention discloses a method for two-dimensional electrophoresis extraction and separation of total protein of a rat and mouse testicular tissue sample. The method comprises rat and mouse testicular tissue treatment, a preparation method for a testicular tissue total protein sample, a two-dimensional electrophoresis technology system, gel dyeing and spectrum analysis. The method optimizes a tedious universal two-dimensional electrophoresis technology system with relatively poor stability, greatly reduces time and reagent consumption for finding out conditions, saves experiment cost, and obtains a satisfactory two-dimensional electrophoresis spectrum that is convenient for subsequent analysis. The method is more suitable for the extraction of the rat and mouse testicular tissue protein sample than a universal homogenate extraction method, a lysate extraction method, a lysate extraction/acetone precipitation method at present. The method not only reduces the degradation of the protein but also increases solubility of the protein without increasing cost of the reagent, so that the two-dimensional electrophoresis spectrum with relatively many protein sites, clear horizontal stripes and relatively good repeatability can be obtained.

Description

technical field [0001] The invention relates to a method for protein extraction and proteomics separation, in particular to a method for whole protein extraction and separation of testicular tissue samples of rats and mice suitable for two-dimensional electrophoresis. Background technique [0002] Protein is the main macromolecule that constitutes the life activities of organisms. With the deciphering of the whole genome sequence of a large number of organisms such as humans and animals and the development of functional genome research, life scientists are paying more and more attention to how to use the genome research model to carry out proteomics research. Proteomics will become one of the strategic commanding heights in the battle for human genes, the largest strategic resource in the new century. At present, proteome research is progressing very rapidly in the world, and both basic theory and technical methods are constantly improving and improving. Since proteomics r...

Claims

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Application Information

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IPC IPC(8): G01N1/34
Inventor 张建海罗广营牛瑞燕王金明孙子龙王俊东
Owner SHANXI AGRI UNIV
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