Application of 4h-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide and its derivatives in glycoprotein-specific fluorescent prestaining detection

Abstract

The present invention relates to a specific fluorescent pre-dyeing detection method detection technology of glycoproteins, particularly to applications of 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide and a derivative thereof in a specific fluorescent pre-dyeing detection method of glycoproteins. The invention further provides a method for using 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide to carry out specific fluorescent pre-dyeing detection of glycoproteins, wherein the method comprises: adding a periodic acid solution to a protein sample to oxidize, adding an ascorbic acid solution to neutralize the excess periodic acid solution, adding a dye to carry out a reaction, adding a loading buffer, carrying out electrophoresis, and observing. According to the present invention, the pre-dyeing has advantages of high sensitivity, good selectivity, simple and rapid operation, good reproducibility, good linearity, good MS compatibility, use safety, low cost, and the like; and compared with the after-gel electrophoresis glycoprotein dyeing method, the pre-dyeing detection method has the following characteristics that the observation can be performed after completing the electrophoresis so as to eliminate the dyeing step after the gel electrophoresis, and the method can be well suitable for the high-throughput proteomics research.

Description

technical field [0001] The invention relates to glycoprotein-specific fluorescent pre-staining detection technology, specifically 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide and its derivatives in glycoprotein-specific Application of fluorescent prestaining assays. Background technique [0002] Protein glycosylation plays a vital role in the field of life science research. As one of the most important and common post-translational modifications of proteins, at least half of mammalian proteins are known to be glycosylated. These Glycoproteins are widely distributed in tissues, cells, and body fluids, especially on the surface of cell membranes, body fluids, etc. Glycosylation plays an important role in protein folding, transportation, and positioning, and participates in receptor activation, signal transduction, etc. Guidance, cell attachment and many other important biological processes. Changes in the number of glycoproteins or changes in the structure o...

AI Technical Summary

Problems solved by technology

[0004] At present, kits in the field of glycoprotein detection are mainly concentrated in post-staining, and most products are sold in the market at high value-added prices. The high price is not conducive to the development of basic biotechnology research

In particular, the cost of using existing biological reagents to carry out biological experiments in China remains high, and it is impossible to achieve the common development of economic benefits and experiments

Glycoprotein-specific fluorescent pre-staining detection technology is rarely reported

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