Non-ampholyte free-flow isoelectric focusing electrophoretic separation method

An isoelectric focusing electrophoresis and ampholyte technology, which is applied in the direction of material analysis, material analysis, and measurement device by electromagnetic means, which can solve the problems of affecting the sensitivity of ultraviolet detection and expensive ampholyte solution.

Inactive Publication Date: 2019-01-25
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the ampholyte solution is not only expensive, but also may

Method used

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  • Non-ampholyte free-flow isoelectric focusing electrophoretic separation method
  • Non-ampholyte free-flow isoelectric focusing electrophoretic separation method
  • Non-ampholyte free-flow isoelectric focusing electrophoretic separation method

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Embodiment 1

[0029] see figure 1 Shown is a schematic diagram of the free-flow electrophoresis device used in this application. The device includes: an operating mobile phase driving pump (1) used to drive the operating mobile phase in the operating mobile phase liquid storage bottle (6) into the gas-liquid buffer device (11). Use the air cylinder (10) in the gas-liquid buffer device (11) to remove the air bubbles in the mobile phase, so that the mobile phase is filled with the free flow electrophoresis cavity (12); the negative electrode liquid circulation pump (2) is used to drive the negative electrode The negative electrode liquid circulation in the liquid storage tank (5), the positive electrode liquid circulation pump (3) is used to drive the positive electrode liquid circulation in the positive electrode liquid storage tank (8); a sample pump (4) is used for The sample in the sample solution storage bottle (7) is pumped into the free-flow electrophoresis chamber (12); a high-voltag...

Embodiment 2

[0033]The first step is to turn on the operating mobile phase to drive the pump (1) to pump the operating mobile phase in the operating mobile phase liquid storage bottle (6) into the gas-liquid buffer device (11). Use the gas cylinder (10) in the gas-liquid buffer device (11) to remove the air bubbles in the running mobile phase, make the running mobile phase fill the free-flow electrophoresis cavity (12) and control the flow rate at 1ml / min. Turn on the negative electrode liquid circulation pump (2) and the positive electrode liquid circulation pump (3) to pump the positive and negative electrode liquids in the negative electrode liquid storage tank (5) and the positive electrode liquid storage tank (8) into the free flow electrophoresis chamber In the positive and negative electrode chambers, and maintain the circulation of the electrode solution. Turn on the high voltage power supply (9) and adjust to 400V voltage. Keep the device running stably for 30 minutes to make it ...

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Abstract

The invention discloses a non-ampholyte free-flow isoelectric focusing electrophoretic separation method. The method comprises the steps that 1, a driving pump is used for pumping an operation flowingphase into a separation cavity of a free-flow electrophoretic device and keeping stable operation of the flow phase; 2, an electrode liquid circulation pump is used for pumping positive electrode liquid in an electrode liquid storage tank into a positive electrode chamber of the free-flow electrophoretic device and keeping circulation of the electrode liquid and used for pumping negative electrode liquid in the electrode liquid storage tank to a negative electrode chamber of the free-flow electrophoretic device and keeping circulation of the electrode liquid; 3, voltage is applied to two electrodes and adjusted to the proper value, stable operation of the device is kept, so that the device is located at the rear portion of the separation cavity, and the stable pH gradient is automaticallyformed between the electrodes. The method has the advantages that the method is simple and easy to implement, the price of the required reagent is low, and sample detection after separation cannot beinfluenced.

Description

technical field [0001] The invention relates to the technical field of free-flow electrophoresis separation, in particular to an ampholyte-free free-flow isoelectric focusing electrophoresis separation method. Background technique [0002] Protein is the material basis of life activities, with a wide variety and diverse functions. It is a biomacromolecule with specific structure and function, which is composed of amino acids linked by amide bonds. It can perform a variety of biological functions, making the information contained in DNA visible. Protein is the most direct embodiment of life activities, and only the study of proteome can fundamentally explain the life process. Protein separation technology is the basis of proteome research. However, due to the special properties of protein itself, it is difficult to use conventional separation techniques for effective separation. [0003] Free-flow electrophoresis is a continuous all-liquid phase separation technique, and ...

Claims

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Application Information

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IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 张凌怡王帅许森张维冰
Owner EAST CHINA UNIV OF SCI & TECH
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