Coomassie brilliant blue staining method, related fixative and related staining agent

Abstract

A novel coomassie brilliant blue staining method, a related fixative and a related staining agent are disclosed. The related fixative comprises an acid and an alcohol, wherein the alcohol is methanol and / or ethanol, and the acid is an acid mixture of acetic acid phosphoric acid in a volume ratio of 1:1. The related staining agent comprises 0.05-0.12% by mass / volume concentration of coomassie brilliant blue, 5-20 v% of an alcohol, 3-10 v% of an acid and 3-10 % by mass / volume concentration of ammonium sulfate. The method is characterized by better linearity, better mass spectrum compatibility, breakage-proof colloid, little background, reduction of nearly a half of agent using amounts, reduced staining cost, and capability of being more environmental friendly.

Description

Technical field [0001] The present invention relates to the field of gel electrophoresis, and more specifically to related reagents and methods for Coomassie brilliant blue staining used in gel electrophoresis. Background technique [0002] Gel electrophoresis techniques, especially one-way SDS-PAGE gel electrophoresis (1-DE) and two-dimensional gel electrophoresis (2-DE) are currently the most widely used protein separation techniques in proteomics research. The basis of quantification is to dye the protein on the gel with a specific dye, and then quantify the protein based on the intensity of the dye. Therefore, the sensitivity, linear range and quantification accuracy based on gel quantification are highly dependent on the level of protein staining. The existing dyeing methods are mainly divided into four types: organic dye dyeing, negative dyeing, silver dyeing and fluorescent dye dyeing. Among them, organic reagent staining is represented by Coomasie brilliant blue (CBB), ...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a kind of economical and environment-friendly Coomassie brilliant blue gel staining method and its corresponding gel fixative and staining agent applied in the field of proteomics research, aiming to form a new staining method, reduce the staining reagent Consumption, solve the problem of difficult decolorization and have high dyeing sensitivity, with significant economic and environmental protection characteristics

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