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Application of Eosin and Its Derivatives in Protein Detection

A protein detection and protein technology, which is applied in the preparation of test samples, color/spectral characteristic measurement, etc., can solve the problems of poor mass spectrometry compatibility, poor reproducibility, and high price, and achieve good reproducibility, low cost, and environmental protection. The effect of little pollution

Inactive Publication Date: 2016-08-31
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

Although many new staining techniques have been developed in the past 30 to 40 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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  • Application of Eosin and Its Derivatives in Protein Detection
  • Application of Eosin and Its Derivatives in Protein Detection
  • Application of Eosin and Its Derivatives in Protein Detection

Examples

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Embodiment 1

[0025] Example 1 Negative staining of eosin protein

[0026] figure 1 is the chemical structural formula of eosin.

[0027] Figure 2-4 The eosin protein negative staining experiment was performed using the following steps:

[0028] 1) Fix the protein sample (SDS6H2) gel after SDS-PAGE electrophoresis in 50% methanol / 10% acetic acid solution for 30 min. Then wash with deionized water twice, each time for 5 minutes;

[0029] 2) Equilibrate in 50% methanol solution for 10 minutes;

[0030] 3) Dye in the staining solution for 15 minutes, then rinse in deionized water for 3 minutes. The staining solution is an aqueous solution containing 0.5% (W / V) eosin and 50% methanol / 1% acetic acid;

[0031] 4) After the gel was stained, it was scanned under the Epson V700 scanner in the reverse scanning mode.

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Abstract

The invention relates to protein negative staining detection technology, in particular to the application of eosin and its derivatives in protein negative staining detection. The invention also provides a method for protein negative staining detection using eosin, comprising the steps of: fixing the gel after protein sample electrophoresis in a fixative solution, washing with deionized water; equilibrating with methanol solution, staining; and developing. The invention has the advantages of high sensitivity, simple and rapid operation, good reproducibility, good linear relationship, good mass spectrometry compatibility, safe use, low cost, etc., and can be better applied to the research of high-throughput 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. The current state-of-the-art 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 the past 30 to 40 years, there are still one or ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/25G01N1/30
Inventor 金利泰丛维涛
Owner 温州安得森生物科技有限公司
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