Polyacrylamide gel and kit thereof

Abstract

The invention relates to a polyacrylamide gel. The polyacrylamide gel is prepared from a separation gel and a concentration gel, wherein the separation gel is prepared from 6 to 15wt% of polyacrylamide, 0.35 to 0.4M of tris(hydroxymethyl)methyl aminomethane, 0.03 to 0.2wt% of tetraethylethylenediamine, 0.1 to 1wt% of sodium hydrogen sulfite and 0.08 to 0.15wt% of ammonium persulfate; the concentration gel is prepared from 4 to 5wt% of polyacrylamide, 0.1 to 0.15M of tris(hydroxymethyl)methyl aminomethane, 0.08 to 0.4wt% of tetraethylethylenediamine, 0.05 to 0.5wt% of sodium hydrogen sulfite and 0.08 to 0.15wt% of ammonium persulfate. The polyacrylamide gel has the advantages that under the synergistic function of multiple components, the stable setting speed of the gel is ensured, and thelong-term stability of the gel is improved; the effect of protein separating by gel electrophoresis is good, and the protein stripes are clear; by not adding the volatile poisonous accelerant TEMED, the injury to the experiment person by the volatile poisonous compound is avoided.

Description

technical field [0001] The invention belongs to polyacrylamide gel electrophoresis, in particular to a polyacrylamide gel and a kit thereof. Background technique [0002] Polyacrylamide gel electrophoresis (PAGE) is the most widely used and most basic experiment in biological experiments. It has two forms: non-denaturing polyacrylamide gel electrophoresis (Native-PAGE) and polyacrylamide gel electrophoresis (PAGE). Acrylamide gel (SDS-PAGE). At present, the traditional PAGE is mainly composed of monomer acrylamide and methylene bisacrylamide with ammonium persulfate (APS) as the catalyst and tetramethylethylenediamine (TEMED) as the accelerator. During the polymerization process, TEMED catalyzes Ammonium persulfate generates free radicals, which initiate the polymerization of acrylamide monomers, and at the same time, methylene bisacrylamide and acrylamide chains generate methylene bond crosslinks, thereby forming a three-dimensional network structure and making a gel. How...

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Problems solved by technology

[0003] 1. TEMED is volatile, flammable, corrosive, and has strong neurotoxicity and irritating odor

[0004] 2. The preparation process is complicated, and various solutions need to be mixed

The glue making process is time-consuming, which greatly reduces the experimental efficiency of the experimenters, and the strong pungent smell of TEMED added later will have a bad impact on the health of the experimenters

[0005] 3. The gel speed is slow. The gel speed of traditional separating gel is 30min-1h, and that of stacking gel is about 30min.

[0006] 4. The stacking gel is colorless, and it is easy to add the sample to the outside of the sample hole when loading the sample, and it is easy to cause contamination between the samples, which will affect the result judgment after protein separation

[0007] At present, there are similar rapid preparation reagents for PAGE gels on the market, but in addition, various adverse experiences occur, such as the need to add TEMED, the electrophoresis speed drops significantly, and the acrylamide solution is easily hydrolyzed and has poor stability, etc.

[0008] The applicant also tried to pre-add TEMED into the separation gel liquid, but this would lead to unstable performance of the solution, easy to decompose, and lose its effect within 3 to 4 days

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