Dyeing and decoloration method of two-dimensional electrophoresis gel Coomassie brilliant blue for screening saline-alkaline resistant plant material

Abstract

The invention discloses a dyeing and decoloration method of two-dimensinal electrophoresis gel Coomassie brilliant blue for screening a saline-alkaline resistant plant material. The method comprises the first step of preparing a staining solution containing Coomassie brilliant blueR250, methyl alcohol, acetic acid and distilled water with a constant volume, uniformly blending the staining solution and still standing for 0.5-1.5 hours, filtering the staining solution and then storing the staining solution at a normal temperature for use; the second step of preparing a destaining solution containing ethyl alcohol, acetic acid and distilled water with a constant volume; the third step of using distilled water to wash two-dimensional SDS gel for 2-3 times, putting the two-dimensional SDS gel in a dyeing box, adding the staining solution into the dyeing box, using a preservative film to seal the dyeing box, and conducting dyeing on the two-dimensional SDS gel on a table concentrator for 2-3 hours at the normal temperature; the fourth step of conducting decoloration, wherein the decoloration comprises the procedures of pouring out the staining solution, using the distilled water to wash the gel for 2-4 times, adding the destaining solution, wherein the dosage of the destaining solution is 1-2 times that of the staining solution, conducting decoloration on the gel on the table concentrator for 2-3 hours at the normal temperature, and using the distilled water to wash the gel for 2-4 times. According to the dyeing and decoloration method, the method is further optimized on the basis of a routine technique method, procedures of pre-immobilization and the like are omitted, the speed of the dyeing and the decoloration is fast, and the effects of the dyeing and the decoloration are good.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a method for rapidly staining and decolorizing a two-dimensional electrophoresis gel by using Coomassie brilliant blue R250. Background technique [0002] Since the function of a gene is mainly realized by its encoded protein, the use of proteomic technology to interpret gene function is considered to be the most important part of post-genome research. At present, the classic proteomics research method is to prepare protein samples, perform two-dimensional electrophoresis, and then obtain a clear protein map by staining, and then obtain the target protein through cutting comparison, enzyme digestion, mass spectrometry and other techniques. In this process, according to the nature of the target protein, different detection methods suitable for SDS gels can be used to detect two-dimensional gels. In order to reduce protein loss as much as possible and improve detection sens...

AI Technical Summary

Problems solved by technology

Silver staining has high sensitivity, but it is prone to high background, resulting in loss of protein resolution, and it is time-consuming, laborious, expensive and requires more exposure to toxic chemicals

Although fluorescent staining has high sensitivity, low background, and mass spectrometry matching, it requires a special scanner and fluorescent dyes, so there is a problem of high staining costs

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