Protein electrophoresis method

Abstract

The invention provides a protein electrophoresis method and relates to the field of protein analysis. The invention particularly relates to preparation of single-layer polyacrylamide gel and a protein electrophoresis analytic method therewith. The preparation of the single-layer polyacrylamide gel is simple in processes and is low in time requirement. The method is excellent in a separation effect on proteins being 4-100 kDa in molecular weight.

Description

technical field [0001] The invention relates to the field of protein analysis, in particular to an electrophoresis method for protein analysis. Background technique [0002] Polyacrylamide gel is formed by acrylamide monomer and a small amount of crosslinker bismethylene acrylamide through chemical catalyst (ammonium persulfate, AP), tetramethylethylenediamine (TEMED) as accelerator or photocatalytic polymerization Polymers in three dimensions. Polyacrylamide gel electrophoresis (PAGE) is a commonly used electrophoresis technique using polyacrylamide gel as a support medium, and is often used for the separation of proteins and nucleic acids. [0003] In denaturing gel electrophoresis, under the action of mercaptoethanol and sodium dodecyl sulfate (SDS), the disulfide bond in the molecule is reduced, and the hydrogen bond is opened to form a SDS-protein complex (SDS and protein mass ratio 1.4 : 1), the complex is negatively charged, so it can migrate to the positive electro...

AI Technical Summary

Problems solved by technology

Conventional SDS-PAGE, corresponding to the effective separation range of proteins is 10~100 kDa (resolving gel concentration is 7.5%~15%), and Tricine-SDS-PAGE, which is commonly used to separate small molecule peptides, corresponds to an effective separation range of 1 ~10 kDa, usually the electrophoresis gel used to separate 1~5 kDa small peptides is composed of three layers of stacking gel, gel layer gel and separating gel, which is relatively cumbersome to make

[0005] When the protein size to be analyzed is in the range of 1-100 kDa, due to the lack of an effective electrophoresis system, it is necessary to perform electrophoresis analysis on SDS-PAGE and Tricine-SDS-PAGE respectively, but the electrophoresis results are produced in different systems. Yes, the two electrophoresis results cannot be effectively compared horizontally, which causes great inconvenience to the experiment

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