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SDS-PAGE separation method for high-and-low-molecular-weight glutenin subunits in wheat

A technology of glutenin and separation method, which is applied in the direction of measuring device, analysis material, material analysis by electromagnetic means, etc., can solve the problems of large amount of medicine, cumbersome operation, long time consumption, etc., and achieves reduction of dosage, shortening of preparation time, The effect of increasing the final concentration

Inactive Publication Date: 2018-11-16
HEBEI ACADEMY OF AGRI & FORESTRY SCI INST OF GENETICS & PHYSIOLOGY
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  • Description
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Problems solved by technology

[0003] Conventional glutenin SDS-PAGE separation method is cumbersome to operate, takes a long time, and consumes a lot of drugs

Method used

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  • SDS-PAGE separation method for high-and-low-molecular-weight glutenin subunits in wheat
  • SDS-PAGE separation method for high-and-low-molecular-weight glutenin subunits in wheat
  • SDS-PAGE separation method for high-and-low-molecular-weight glutenin subunits in wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0048] The operation process is the same as the comparison example, the difference is:

[0049] 1. In the process of gliadin removal and glutenin reduction, reduce the concentration of isopropanol from 50% to 30%, that is, replace extract A1 (50% isopropanol) with extract A2 ( 30% isopropanol), and replace the extract solution B7 with the extract solution B5.

[0050] 2. In the alkylation process of glutenin subunits, after the reduction of glutenin, the alkylation agent 4-VP is directly added to the sample buffer C, and the glutenin subunit extract and the sample Alkylation is carried out during the mixing process of buffer C. The concentration of 4-VP in sample buffer C is 1.0%-0.6%. In the sample buffer C of VP, vortex and mix, treat in a water bath at 65°C for 15 minutes, centrifuge at 10,000g for 2 minutes, and take 10 ul of the supernatant for the next step of SDS-PAGE electrophoresis.

[0051] Specifically, replace steps (3) and (4) with:

[0052] Take 50 ul of the s...

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Abstract

The invention relates to an SDS-PAGE separation method for high-and-low-molecular-weight glutenin subunits in wheat. The SDS-PAGE separation method comprises the following steps: removal of alcohol-soluble proteins; reduction of glutenin; mixed alkylation of glutenin subunits and a sample buffer; and SDS-PAGE. The method of the invention reduces the usage amount of a reagent, omits the preparationof a sample and shortens sample preparation time on the premise of maintaining sample separation effect.

Description

technical field [0001] The invention relates to a SDS-PAGE separation method of wheat high and low molecular weight glutenin subunits in the field of wheat processing quality improvement. Background technique [0002] Wheat glutenins include high molecular weight glutenin (HMW-GS) and low molecular weight glutenin (LMW-GS), which together with gliadin constitute wheat gluten. In the gluten network, different glutenins play different roles. Function, high molecular weight glutenin is the main component of gluten network, it is the skeleton of gluten network, low molecular weight glutenin is connected with high molecular weight glutenin to form a chain structure with fine branches, the composition of glutenin directly affects the strength of wheat gluten, It is an important parameter affecting the quality of wheat processing, and SDS-PAGE is a common method for analyzing the composition of wheat glutenin. [0003] The conventional SDS-PAGE separation method for glutenin is cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 柴建芳李春燕马秀英王海波周硕赵和吕孟雨董福双刘永伟杨帆
Owner HEBEI ACADEMY OF AGRI & FORESTRY SCI INST OF GENETICS & PHYSIOLOGY
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