Two-dimensional gel electrophoresis technology and mass spectrometry technology are currently the most widely used methods for studying proteomics. Two-dimensional gel electrophoresis technology uses protein isoelectric point and molecular weight difference to distinguish various proteins. Although it is difficult to distinguish low-abundance proteins by two-dimensional gel electrophoresis, it has higher requirements for operation, but its high throughput, good resolution and reproducibility, and its characteristics of being combined with mass spectrometry make it the most popular and reliable Proteomics research methods. Two-dimensional gel electrophoresis technology and mass spectrometry-based proteomics research procedures are sample preparation→isoelectric focusing→polyacrylamide gel electrophoresis→gel staining→digging out protein spots of interest→in-gel digestion→mass spectrometry analysis to determine peptides Fingerprint or partial amino acid sequence→use database to determine protein. Proteomics research requires high-resolution protein separation and accurate and sensitive mass spectrometry identification techniques. The coloring of proteins in gel electrophoresis not only affects the resolution of protein separation, but also affects subsequent mass spectrometry identification. Protein staining can be divided into four categories: organic reagent staining, silver staining, fluorescent staining and isotope coloration.
Unlu et al. proposed a fluorescence differential display two-dimensional electrophoresis (F-2D-DIGE) quantitative proteomics analysis method. Differential gel electrophoresis (DIGE) is a technical improvement of 2-DE. It combines the method of multiple fluorescence analysis. It separates multiple samples labeled with different fluorescence on the same gel and introduces the internal The underlying concept. The proteins in the two samples are mixed with different fluorescent labels to perform 2-DE to detect the expression of the protein in the two samples, which greatly improves the accuracy, reliability and repeatability of the results. In DIGE technology, each protein spot has its own internal standard, and the software can automatically calibrate its expression according to the internal standard of each protein spot to ensure that the detected protein abundance changes are true. DIGE technology has been applied in various samples.