Protein isoelectric focusing electrophoresis method and glue preparation device

An isoelectric focusing electrophoresis, protein technology, applied in measuring devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of long electrophoresis time, protein diffusion tailing, etc., to reduce reagent consumption, good separation, Easy-to-use effects

Active Publication Date: 2018-03-30
LIVZON GROUP LIVZON PHARMA FACTORY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In order to overcome the deficiencies in the prior art, the object of the present invention is to provide a protein isoelectric focusing electrophoresis method and a gel dispensing device thereof. The protein electrophoresis strips obtained by this method are clear, have good separation, do not diffuse and do not tail, and are effective It solves the problems of long electrophoresis time and protein diffusion and tailing in the prior art, and greatly improves the success rate of gel dispensing

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  • Protein isoelectric focusing electrophoresis method and glue preparation device
  • Protein isoelectric focusing electrophoresis method and glue preparation device
  • Protein isoelectric focusing electrophoresis method and glue preparation device

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Embodiment

[0043] 1. Instruments and reagents:

[0044] 1. Reagents used: The following reagents are prepared with analytically pure and ultrapure water (resistivity should not be lower than 18MΩ.cm) unless otherwise specified.

[0045] 30% acrylamide solution, ampholyte (PH3-10) solution (purchased by BIO-RAD company), 10% ammonium persulfate solution, N,N,N',N'-tetramethylethylenediamine (TEMED);

[0046] Catholyte (0.5mol / L phosphoric acid solution): take 3.41ml of phosphoric acid, add water to dilute to 100ml and mix well;

[0047] Anode solution (0.2mol / L sodium hydroxide solution): Weigh 0.8g of sodium hydroxide, add water to dissolve it to 100ml and mix well;

[0048] Fixative solution: 34.5g trichloroacetic acid, 10.4g sulfosalicylic acid, add water to dissolve to 300ml.

[0049] Decolorization solution: mix 500ml of 95% ethanol with 160ml of glacial acetic acid, add water to dilute to 2000ml;

[0050] Dyeing solution: 0.35g Coomassie brilliant blue G250 (R250), add 300ml deco...

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Abstract

The invention discloses a protein isoelectric focusing electrophoresis method and glue preparation device, and belongs to the technical field of bioseparation detection. The method comprises the stepsof glue preparation, glue filling, pre-electrophoresis, electrophoresis, gel fixing, decoloration, dyeing and the like. According to the method, the feeding mode of a sample and voltage and electrophoresis time of pre-electrophoresis and electrophoresis are changed, so that obtained protein electrophoresis bands are clear, good in separation degree and do not diffuse or trail, the problem that inthe prior art, the electrophoresis time is long, and protein diffuses and trails is effectively solved, and the glue preparation success rate is greatly increased. The glue preparation device comprises a glue preparation support, a fixed frame, glue preparation glass, a gasket, an anti-leakage pad and other components. By improving the structure of the glue preparation device, the consumption ofa reagent is reduced, glue filling is fast, no bubble is generated, glue gathering is even, the glue preparation success rate is greatly increased, and meanwhile the inspection cost is reduced.

Description

technical field [0001] The invention relates to the technical field of biological separation and detection, in particular to a protein isoelectric focusing electrophoresis method and a glue dispensing device. Background technique [0002] Isoelectric focusing electrophoresis (IEF) is a pH gradient formed by ampholytes in an electrophoresis field (usually polyacrylamide gels). Since proteins are amphoteric substances, their charges are related to the pH of the medium. Migrate in the opposite direction of polarity, when the migration reaches the pH of its isoelectric point (PI), at this point, the protein no longer has a net positive or negative charge, the current reaches a minimum, and no longer moves, forming a very Narrow zone, so as to achieve the electrophoresis method to detect the isoelectric point of protein and peptide test samples. At present, the conventional isoelectric focusing electrophoresis method has the following problems: 1. IEF electrophoresis often cause...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCG01N27/44704G01N27/44795
Inventor 曾娟梅孙朗扈会平陈清华徐晓吴菊莲孙莉钟贞曹珺
Owner LIVZON GROUP LIVZON PHARMA FACTORY
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