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4', 5'-dibromo fluorescein and application of its derivative in protein detection

A dibromofluorescein and protein detection technology, which is applied in the field of protein negative staining detection, can solve the problems of poor mass spectrometry compatibility, cumbersome operation, and low sensitivity, and achieve good reproducibility, simple and rapid operation, and less environmental pollution.

Inactive Publication Date: 2017-05-10
温州安得森生物科技有限公司
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  • Abstract
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Problems solved by technology

Although many new staining techniques have been developed in recent years, there are still one or several disadvantages as follows: low sensitivity, poor reproducibility, high toxicity, poor compatibility with mass spectrometry, high price, cumbersome operation, etc.

Method used

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  • 4', 5'-dibromo fluorescein and application of its derivative in protein detection
  • 4', 5'-dibromo fluorescein and application of its derivative in protein detection
  • 4', 5'-dibromo fluorescein and application of its derivative in protein detection

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Embodiment 14

[0025] Example 1 4′,5′-Dibromofluorescein Negative Staining

[0026] figure 1 is the chemical structural formula of 4',5'-dibromofluorescein.

[0027] Figure 2~4 The 4′,5′-dibromofluorescein protein negative staining experiment was performed using the following steps:

[0028] 1) The gel after electrophoresis was stained in 0.3% 4′,5′-dibromofluorescein / 40% methanol / 10% glycerol / 50 μM zinc chloride staining solution for 8 minutes.

[0029] 2) The staining solution was discarded, and developed in 100 mM sodium acetate-acetic acid buffer solution (pH=4.6) for 2 min.

[0030] 3) After gel staining, scan in reverse scanning mode on Epson V700 scanner.

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Abstract

The invention relates to a protein negative staining detection technology, and particularly relates to 4', 5'-dibromo fluorescein and application of its derivative in protein detection. The invention further provides a method of performing protein negative staining detection by applying the 4', 5'-dibromo fluorescein. The method includes steps of directly dyeing after finishing the protein gel electrophoresis, and then developing. The 4', 5'-dibromo fluorescein has advantages of high sensitivity, simple operation, good reproducibility, good mass spectrum compatibility, low cost, and others, and can be well applicable to the study of high-flux proteomics.

Description

technical field [0001] The invention relates to protein negative stain detection technology, in particular to a new negative dye for protein detection. Background technique [0002] In proteomics research, gel protein staining technology is a key technology connecting two-dimensional electrophoresis and downstream mass spectrometry analysis. Current advanced mass spectrometers are capable of analyzing trace proteins at the pg level. Conventional gel protein staining techniques include Coomassie brilliant blue staining, silver staining, and fluorescent staining. The Coomassie Brilliant Blue staining method has low sensitivity, the silver staining method is complicated to operate, has poor compatibility with mass spectrometry, and the fluorescent staining method is expensive. These factors seriously restrict the development of high-throughput proteomics. Although many new staining techniques have been developed in recent years, there are still one or several disadvantages a...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N33/68
CPCG01N1/30G01N33/6848G01N2001/302
Inventor 金利泰
Owner 温州安得森生物科技有限公司
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