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Method for in situ fluorescence derivation in protein membrane

A protein and fluorescence technology, applied in the field of protein fluorescence detection, can solve the problems of poor fluorescence derivatization effect and poor reaction effect of low-concentration protein, and achieve the effect of improving fluorescence detection effect, simple device and good derivatization effect.

Inactive Publication Date: 2009-06-24
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology

However, the method of enriching proteins after the derivatization reaction cannot fundamentally solve the problem of poor fluorescence derivatization effect of low-concentration proteins
When the protein concentration is very low, the protein reacts poorly with fluorescent reagents, so only relying on enrichment after derivatization cannot fundamentally solve the problem

Method used

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  • Method for in situ fluorescence derivation in protein membrane
  • Method for in situ fluorescence derivation in protein membrane
  • Method for in situ fluorescence derivation in protein membrane

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Embodiment

[0021] see figure 1 and figure 2 , the method of in situ fluorescence derivation of protein membrane includes two steps: enrichment of protein in hollow fiber membrane; fluorescence derivation of protein after enrichment. The components required for the protein enrichment process include a pump 1 , a capillary 2 , a cellulose acetate hollow fiber membrane 3 , and a rubber sealing head 4 . The protein sample is driven by the pump 1 and continuously flows into the hollow fiber membrane 3 at a certain flow rate. Since the outlet of the hollow fiber membrane is sealed by the rubber head 4, the protein can be enriched in the membrane. After enrichment of protein samples, fluorescence derivatization is performed. The hollow fiber membrane enriched with the protein sample is spirally put into the centrifuge tube 5, and then the fluorescent reagent 6 is added into the centrifuge tube, and the hollow fiber membrane 3 is submerged. The tubes are heated and stirred magnetically for p...

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Abstract

The invention relates to protein fluorescence detection, in particular to a method for in-vivo fluorescence derivation in a protein film, which comprises the steps as follows: firstly, a protein solution is pumped from one end of a cellulose acetate hollow fiber membrane (3) into the cellulose acetate hollow fiber membrane (3) through a pump (1), and the other end thereof is used as an outlet that is closed; and the outlet of the hollow fiber membrane (3) is shut, so that proteins can be enriched in the membrane; and secondly, the hollow fiber membrane enriched with protein samples is helically put in a centrifugal tube (5), then a fluorescent reagent solution (6) is added in the centrifugal tube, and the hollow fiber membrane (3) is submerged; the centrifugal tube is heated and processed through magetic stirring for the derivation of the proteins. The method has the advantages of simple equipment, easy operation and low cost, and can perform fluorescence derivation to light concentration proteins.

Description

technical field [0001] The invention relates to the fluorescence detection of protein, in particular to a method for in-situ fluorescence derivation in protein membranes. Proteins are enriched in hollow fiber membranes, and then the hollow fiber membranes are immersed in fluorescent reagents for protein derivation. Background technique [0002] Fluorescence detection has extremely high detection sensitivity. With the introduction of proteomics, many articles have applied fluorescence-derived methods to the detection of proteins. Through fluorescence derivatization, the detection sensitivity of proteins has been greatly improved. However, there are problems during the fluorescent derivatization of proteins at lower concentrations. Since the derivatization reaction of the protein is related to the concentration, when the protein concentration is very low, the derivatization effect is poor, which affects the fluorescence detection process. Banks et al. [Lau, S.K.; Zaccardo, ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/00
Inventor 张丽华乔晓强孙良亮梁玉张维冰梁振张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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