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

A fluorescence detection and glycoprotein technology, applied in the field of glycoprotein-specific fluorescence detection, can solve the problems of inability to realize the common development of economic benefits and experiments, unfavorable development of basic biotechnology research, high cost of biological experiments, etc. Simple and fast operation, less environmental pollution

Inactive Publication Date: 2017-08-01
温州安得森生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, kits in the field of glycoprotein detection are mainly concentrated in foreign biological reagent companies, and most of the 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

Method used

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  • Application of 4h-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide and its derivatives in specific fluorescence detection of glycoproteins
  • Application of 4h-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide and its derivatives in specific fluorescence detection of glycoproteins
  • Application of 4h-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide and its derivatives in specific fluorescence detection of glycoproteins

Examples

Experimental program
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Effect test

experiment example 14

[0028] Experimental example 1 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein-specific fluorescent staining

[0029] figure 1 It is the chemical structural formula of 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide.

[0030] 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein fluorescent staining method is carried out as follows:

[0031] 1) Fix the protein sample gel after SDS-PAGE electrophoresis in 40% ethanol and 10% acetic acid aqueous solution for 30min, discard the fixative;

[0032] 2) Oxidation in a periodate solution for 20 minutes, wherein the periodate solution is an aqueous solution containing 0.5% by volume of periodic acid and 1% by volume of acetic acid. Then rinse with 1% acetic acid aqueous solution by volume for 3 times, each time for 5 minutes;

[0033] 3) Add dyeing solution to dye for 10 minutes, wherein the dyeing solution is 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide c...

experiment example 24

[0037] Experimental example 2 Comparison of detection effect of standard protein by 4H-[1]-benzopyran[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein fluorescent staining method and other staining methods.

[0038] (A) 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein fluorescent staining method, (B) Pro-QEmerald glycoprotein fluorescent staining method, (C) SYPRO Ruby Whole protein fluorescent staining. Both the Pro-Q Emerald glycoprotein fluorescent staining method and the SYPRO Ruby whole protein fluorescent staining method were described in the literature; 8 different standard proteins from Sigma were used as samples. Band 1, 1000ng; Band 2, 500ng; Band 3, 250ng; Band 4, 125ng; Band 5, 64ng; Band 6, 32ng; Band 7, 16ng; Band 8, 8ng; Band 9, 4ng; Band 10, 2ng. The result is as figure 2 As shown, it shows that the detection sensitivity of 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein fluorescence staining met...

experiment example 34

[0039] Experimental Example 3 Comparison of the detection effect of 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein fluorescent staining method and other staining methods on human serum total protein.

[0040] (A) 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein fluorescent staining method, (B) Pro-QEmerald glycoprotein fluorescent staining method, (C) SYPRO Ruby Whole protein fluorescent staining. The Pro-Q Emerald glycoprotein fluorescent staining method and the SYPRO Ruby whole protein fluorescent staining method were operated according to the literature; the extracted human serum total protein was used as the sample. Belt 1, 5000ng; The result is as image 3 As shown, it shows that the detection sensitivity of 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide glycoprotein fluorescence staining method is close to that of Pro-Q Emerald488 glycoprotein fluorescence staining method.

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Abstract

The invention relates to glycoprotein specific fluorescence detection technology, and particularly relates to application of 4 H-[1]-benzo pyran [4, 3-b] thiophene-2-formic acid hydrazide and a derivative thereof in glycoprotein specific fluorescence detection. The invention also provides a method for the application of the 4 H-[1]-benzo pyran [4, 3-b] thiophene-2-formic acid hydrazide for glycoprotein specific fluorescence detection, and the method comprises the steps of: fixation of after-electrophoresis gel containing a protein sample in a fixing solution; oxidation with periodic acid solution; washing with acetic acid aqueous solution; dyeing; and elution. The invention has the advantages of high sensitivity, good selectivity, fast and simple operation, good reproducibility, good linear relationship, good mass spectrum compatibility, safe use, low cost, and the like, and can be better suitable for the study of high-throughput proteomics.

Description

technical field [0001] The present invention relates to glycoprotein-specific fluorescence detection technology, specifically 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide and its derivatives in glycoprotein-specific fluorescence detection in the application. 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 of sugar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 金利泰丛维涛朱忠欣洪国赢
Owner 温州安得森生物科技有限公司
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