A method for separating wheat high and low molecular weight glutenin subunit sds-page
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 cumbersome operation, long time-consuming, high dosage of medicine, etc., and achieves reduction of dosage, shortening of preparation time, The effect of increasing the final concentration
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[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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