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Synthesis of 1-pyrenyl-carbohydrazide and application of 1-pyrenyl-carbohydrazide in specific detection of glycoprotein

A technology of pyrenylcarbohydrazide and glycoprotein, which is applied in the field of glycoprotein-specific fluorescence detection, can solve the problems of unfavorable research and development of glycoproteomics, the inability to realize economic benefits and experiments, and high cost, and achieve simple operation Rapid, low cost, and less environmental pollution effects

Inactive Publication Date: 2015-04-08
温州安得森生物科技有限公司
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  • Abstract
  • Description
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Problems solved by technology

However, most products are expensive and the detection steps are cumbersome, which is not conducive to the development of glycoproteomics research
Moreover, on the basis of existing biological reagents in China, the cost of conducting biological experiments remains high, and it is impossible to achieve the common development of economic benefits and experiments.

Method used

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  • Synthesis of 1-pyrenyl-carbohydrazide and application of 1-pyrenyl-carbohydrazide in specific detection of glycoprotein
  • Synthesis of 1-pyrenyl-carbohydrazide and application of 1-pyrenyl-carbohydrazide in specific detection of glycoprotein
  • Synthesis of 1-pyrenyl-carbohydrazide and application of 1-pyrenyl-carbohydrazide in specific detection of glycoprotein

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Experimental program
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Embodiment 1

[0028] Example 1 UGF202 glycoprotein-specific fluorescent staining

[0029] figure 1 is the chemical structural formula of 1-pyrenylcarbohydrazide.

[0030] The UGF202 glycoprotein fluorescent staining method is carried out as follows:

[0031] 1) Fix the protein sample gel after SDS-PAGE electrophoresis in 50% methanol and 5% acetic acid aqueous solution for 30min, discard the fixative;

[0032] 2) Oxidation in a periodic acid solution for 20 minutes, wherein the periodic acid solution is an aqueous solution containing 0.5% periodic acid by weight and volume and 3% by volume acetic acid. Then wash once with 0.8% vitamin C aqueous solution by weight to volume for 5 minutes;

[0033] 3) adding dyeing solution for dyeing for 20 minutes, wherein the dyeing solution is UGF202 containing 0.0001% by volume ratio of UGF202 and 5% by volume of DMF and 40% ethanol solution;

[0034] 4) Detection, the stained protein can be observed under a laser scanner.

[0035] Derivatives such ...

experiment example 1U

[0036] Experimental example 1 Comparison of the detection effect of UGF202 glycoprotein fluorescent staining method and other staining methods on standard protein.

[0037] (A) UGF202 glycoprotein fluorescent staining method, (B) Pro-Q Emerald 300 glycoprotein fluorescent staining method, (C) Pro-Q Emerald 488 glycoprotein fluorescent staining method, (D) SYPRO Ruby whole protein fluorescent staining method. 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, 250ng; Band 2, 125ng; Band 3, 64ng; Band 4, 32ng; Band 5, 16ng; Band 6, 8ng; Band 7, 4ng; Band 8, 2ng; Band 9, 1ng; Band 10, 0.5ng . The result is as figure 2 As shown, the detection sensitivity of UGF202 glycoprotein fluorescent staining method is slightly better than that of Pro-Q Emerald 300 glycoprotein fluorescent staining method, and is 8...

experiment example 2

[0038]Experimental example 2 Comparison of the detection effect of UGF202 glycoprotein fluorescent staining method and other staining methods on different samples.

[0039] (A) UGF202 glycoprotein fluorescent staining method, (B) Pro-Q Emerald 300 glycoprotein fluorescent staining method, (C) Pro-Q Emerald4 88 glycoprotein fluorescent staining method, (D) SYPRO Ruby whole protein fluorescent staining method, all Operate as documented. The four samples used are: total protein extracted from human serum, total protein extracted from human fibroblasts, total protein extracted from mouse liver tissue, and total protein extracted from mouse myocardium. Among them, the extracted human serum total protein is used as the sample, and the loading amount is: band 1,500ng; band 2,250ng; band 3,125ng; band 4,64ng; band 5,32ng; band 6,16ng; band 7, 8ng; belt 8, 4ng; belt 9, 2ng; belt 10, 1ng. The result is as image 3 As shown, the detection sensitivity of UGF202 glycoprotein fluorescent...

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Abstract

The invention relates to the field of specific fluorescence detection technology of glycoprotein in the research orientation and in particular relates to synthesis of 1-pyrenyl-carbohydrazide (UGF202) and application of derivatives of 1-pyrenyl-carbohydrazide in specific detection of glycoprotein. The invention provides a method for performing specific fluorescence detection on glycoprotein by virtue of 1-pyrenyl-carbohydrazide. The method comprises the following four steps: fixing gel containing a protein sample subjected to electrophoretic separation in a fixing solution; oxidizing with a periodic acid solution; washing with vitamin C aqueous solution; and staining. The method has extremely high sensitivity, can be used for detecting glycoprotein lowering to 0.5ng, has the advantages of high specificity, simple and convenient operation steps, time conservation, high reproducibility, good linear relation, high mass spectrum compatibility, safety in use and low cost and can be well applied to research of high-flux proteomics.

Description

technical field [0001] The invention relates to a glycoprotein-specific fluorescence detection technology, in particular to the design and synthesis of 1-pyrenyl-carbohydrazide (UGF202) and its application in glycoprotein-specific fluorescence detection. Background technique [0002] Glycosylation is one of the most important ways of protein post-translational modification and is an essential physiological process. At present, it is known that at least one-half of mammalian proteins are glycosylated, and these glycoproteins are widely distributed in tissues, cells, and body fluids, especially in cell membrane surfaces, body fluids, etc. Playing it right plays a vital role. For example, the sugar chain part of glycoprotein affects the folding, solubility, half-life, antigenicity and biological activity of the protein; the molecular basis of the interaction between proteins, the specific binding of protein and receptor, etc. Recognition; Many malignant tumor tissues show mar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/64
CPCC07C281/14C09K11/06C09K2211/1011G01N21/6428
Inventor 金利泰
Owner 温州安得森生物科技有限公司
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