Application of Palmatine and its derivatives in protein fluorescent detecting

A protein fluorescence and derivative technology, applied in the field of protein fluorescence detection, to achieve the effects of good reproducibility, less environmental pollution and low cost

Inactive Publication Date: 2008-01-30
WENZHOU MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there have been no reports of its use in protein fluorescent staining

Method used

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  • Application of Palmatine and its derivatives in protein fluorescent detecting
  • Application of Palmatine and its derivatives in protein fluorescent detecting
  • Application of Palmatine and its derivatives in protein fluorescent detecting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Palmatine protein fluorescent staining

[0031] Figure 1 is the chemical structural formula of palmatine. The palmatine protein fluorescent staining experiment was carried out using the following steps:

[0032] 1) The gel of the protein sample (SDS-6H, Sigma-Aldrich Co, USA) after SDS-PAGE electrophoresis was placed in a fixative solution for 5 minutes, rinsed with deionized water for 3 minutes, and the fixative solution was 40% ethanol by volume, 10% acetic acid aqueous solution;

[0033] 2) Dye in the staining solution for 10 minutes, then rinse with deionized water for 0.5 minutes, the staining solution is an aqueous solution containing 0.0005% (W / V) palmatine and 2% acetic acid;

[0034] 3) Put the dyed gel directly on the transilluminator, transmit at 312nm, and record the image.

[0035] According to the above method, the hydrochloride, sulfate or hydroiodide of palmatine were used for protein staining, and the results showed that detection results s...

experiment example 1

[0037] Experimental Example 1 Effect of deionized water washing on sensitivity before gel staining

[0038] Stain according to the method of Example 1, A is the gel obtained without washing before staining, B is the gel rinsed for 3 minutes before staining, the protein (protein style) sample loading (each band) is as follows: band 1, 1000ng; Band 2, 2500ng; Band 3, 250ng; Band 4, 125ng; Band 5, 63ng; Band 6, 32ng; Band 7, 16ng; Band 8, 8ng; Band 9, 4ng; Band 10, 2ng. Explain that washing in step 2 is necessary, which can significantly improve the staining effect.

experiment example 2

[0039] Influence of Experimental Example 2 Palmatine Concentration on Dyeing Effect

[0040] According to the method of embodiment 1, dye with different concentrations of palmatine, the concentration of palmatine is respectively: (a) 0.0001%, (b) 0.0003%, (c) 0.0005%, (d) 0.0007%, ( e) 0.0009%, the results are shown in Fig. 3A. Experiments show that a better dyeing effect can still be obtained when the palmatine concentration is 0.0001%, but the effect is better at 0.0005%. The quality of standard protein loading ranged from 4ng to 1000ng / band, and the brightness of the protein band was analyzed by Tina2.09 image analysis software (the same below).

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Abstract

The invention relates to a protein fluorescence detection technology, in particular to the application of the THP and the derivatives in the protein fluorescence detection. The invention also provides a method to apply the THP to the protein fluorescence gel detection, and the method comprises the steps of: fixing the electrophoresis gel of the protein sample in the stationary liquid, washing by the deionized water; dyeing, washing by the deionized water; detection. The invention has the advantages of high sensitive, simple and rapid operation, good reproducibility, good dyeing background, good linear relationship, good reversibility, good compatibility, safe using and low cost, etc.

Description

technical field [0001] The invention relates to protein fluorescence detection technology, in particular to a new fluorescent dye of protein. Background technique [0002] In the field of life science research, it is inseparable from the research on protein and DNA, the basic substances of life, and also inseparable from the detection technology of protein. Especially in recent years, with the rapid development of genomics and proteomics, we have higher requirements for protein separation and detection technology. [0003] There are many methods for monitoring protein, including turbidimetry, Kjeldahl method, absorbance photometry, fluorescence photometry, and the newly developed resonance Rayleigh scattering method, high performance liquid chromatography, and capillary electrophoresis analysis. Fluorescence photometry is currently the most researched, most widely used, easy-to-operate, and highly sensitive detection method. [0004] Palmatine belongs to the proberberine c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/52G01N21/76C09K11/06
Inventor 金利泰
Owner WENZHOU MEDICAL UNIV
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